Apparatus and method for electromagnetic treatment of neurological injury or condition caused by a stroke

ABSTRACT

Described herein are methods of treating neurological injury and conditions, in particular symptoms and complications arising from or caused by a stroke. These treatment methods can include the steps of generating a pulsed electromagnetic field from a pulsed electromagnetic field source and applying the pulsed electromagnetic field in proximity to a target region.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.14/171,553, filed on Feb. 3, 2014, entitled “APPARATUS AND METHOD FORELECTROMAGNETIC TREATMENT OF NEUROLOGICAL INJURY OR CONDITION CAUSED BYA STROKE,” Publication No. US-2014-0213843-A1, which is acontinuation-in-part of U.S. patent application Ser. No. 13/252,114,filed on Oct. 3, 2011, entitled “METHOD AND APPARATUS FORELECTROMAGNETIC TREATMENT OF HEAD, CEREBRAL AND NEURAL INJURY IN ANIMALSAND HUMANS,” Publication No. US-2012-0116149-A1, which claims thebenefit under 35 U.S.C. §119 of U.S. Provisional Application No.61/389,038 filed on Oct. 1, 2010 and U.S. Provisional Application No.61/456,310 filed on Nov. 4, 2010, the disclosures of which areincorporated by reference as if fully set forth herein.

U.S. patent application Ser. No. 14/171,553 is also acontinuation-in-part of U.S. patent application Ser. No. 13/801,789filed on Mar. 13, 2013, entitled “APPARATUS AND METHOD FORELECTROMAGNETIC TREATMENT,” Publication No. US-2013-0274540-A1, which isa continuation of U.S. patent application Ser. No. 12/819,956, filed onJun. 21, 2010, entitled “APPARATUS AND METHOD FOR ELECTROMAGNETICTREATMENT,” Publication No. US-2011-0112352-A1, now abandoned, which isa continuation-in-part of U.S. patent application Ser. No. 12/772,002,filed on Apr. 30, 2010, entitled “APPARATUS AND METHOD FORELECTROMAGNETIC TREATMENT OF PLANT, ANIMAL AND HUMAN TISSUE, ORGANS,CELLS AND MOLECULES,” Publication No. US-2010-0222631-A1, now abandoned,which is a continuation of U.S. patent application Ser. No. 11/003,108,filed on Dec. 3, 2004, entitled “APPARATUS AND METHOD FORELECTROMAGNETIC TREATMENT OF PLANT, ANIMAL, AND HUMAN TISSUE, ORGANS,CELLS, AND MOLECULES,” now U.S. Pat. No. 7,744,524, which claims thebenefit under 35 U.S.C. §119 of U.S. Provisional Patent Application No.60/527,327, filed on Dec. 5, 2003, entitled “APPARATUS AND METHOD FORELECTROMAGNETIC TREATMENT OF PLANT, ANIMAL, AND HUMAN TISSUE, ORGANS,CELLS AND MOLECULES.”

U.S. patent application Ser. No. 12/819,956 is also acontinuation-in-part of U.S. patent application Ser. No. 11/114,666,filed on Apr. 26, 2005, entitled “ELECTROMAGNETIC TREATMENT INDUCTIONAPPARATUS AND METHOD FOR USING SAME,” now U.S. Pat. No. 7,740,574, whichclaims the benefit under 35 U.S.C. §119 of U.S. Provisional PatentApplication No. 60/564,887, filed on Apr. 26, 2004, entitled “INDUCTIONMEANS FOR THERAPEUTICALLY TREATING HUMAN AND ANIMAL CELLS, TISSUES ANDORGANS WITH ELECTROMAGNETIC FIELDS.”

U.S. patent application Ser. No. 12/819,956 is also acontinuation-in-part of U.S. patent application Ser. No. 11/110,000,filed on Apr. 19, 2005, entitled “ELECTROMAGNETIC TREATMENT APPARATUSAND METHOD FOR ANGIOGENESIS MODULATION OF LIVING TISSUES AND CELLS,”Publication No. US-2005-0251229-A1, now abandoned, which claims thebenefit under 35 U.S.C. §119 of U.S. Provisional Patent Application No.60/563,104, filed on Apr. 19, 2004, entitled “APPARATUS AND METHOD FORTHERAPEUTICALLY TREATING HUMAN AND ANIMAL CELLS, TISSUES AND ORGANS WITHELECTROMAGNETIC FIELDS.”

U.S. patent application Ser. No. 12/819,956 is also acontinuation-in-part of U.S. patent application Ser. No. 11/369,308,filed on Mar. 6, 2006, entitled “ELECTROMAGNETIC TREATMENT APPARATUS FORAUGMENTING WOUND REPAIR AND METHOD FOR USING SAME,” Publication No.US-2006-0212077-A1, now abandoned, which claims the benefit under 35U.S.C. §119 of U.S. Provisional Patent Application No. 60/658,967, filedon Mar. 7, 2005, entitled “APPARATUS AND METHOD FOR THERAPEUTICALLYTREATING HUMAN, ANIMAL, AND PLANT CELLS, TISSUES, ORGANS, AND MOLECULESWITH ELECTROMAGNETIC FIELDS FOR WOUND REPAIR.”

U.S. patent application Ser. No. 12/819,956 is also acontinuation-in-part of U.S. patent application Ser. No. 11/369,309,filed on Mar. 6, 2006, entitled “ELECTROMAGNETIC TREATMENT APPARATUS FORENHANCING PHARMACOLOGICAL, CHEMICAL AND TOPICAL AGENT EFFECTIVENESS ANDMETHOD FOR USING SAME,” Publication No. US-2007-0026514-A1, nowabandoned, which claims the benefit under 35 U.S.C. §119 of U.S.Provisional Patent Application No. 60/658,968, filed on Mar. 7, 2005,entitled “APPARATUS AND METHOD FOR TREATING HUMAN, ANIMAL AND PLANTCELLS, TISSUES, ORGANS AND MOLECULES WITH ELECTROMAGNETIC FIELDS BYENHANCING THE EFFECTS OF PHARMACOLOGICAL, CHEMICAL, COSMETIC AND TOPICALAGENTS.”

U.S. patent application Ser. No. 12/819,956 is also acontinuation-in-part of U.S. patent application Ser. No. 11/223,073,filed on Sep. 10, 2005, entitled “INTEGRATED COIL APPARATUS FORTHERAPEUTICALLY TREATING HUMAN AND ANIMAL CELLS, TISSUES AND ORGANS WITHELECTROMAGNETIC FIELDS AND METHOD FOR USING SAME,” now U.S. Pat. No.7,758,490.

U.S. patent application Ser. No. 12/819,956 is also acontinuation-in-part of U.S. patent application Ser. No. 11/339,204,filed on Jan. 25, 2006, entitled “SELF-CONTAINED ELECTROMAGNETICAPPARATUS FOR TREATMENT OF MOLECULES, CELLS, TISSUES, AND ORGANS WITHINA CEREBROFACIAL AREA AND METHOD FOR USING SAME,” Publication No.US-2007-0173904-A1, now abandoned.

U.S. patent application Ser. No. 12/819,956 is also acontinuation-in-part of U.S. patent application Ser. No. 11/818,065,filed on Jun. 12, 2007, entitled “ELECTROMAGNETIC APPARATUS FORPROPHYLAXIS AND REPAIR OF OPHTHALMIC TISSUE AND METHOD FOR USING SAME,”Publication No. US-2008-0058793-A1, now abandoned, which claims thebenefit under 35 U.S.C. §119 of U.S. Provisional Patent Application No.60/812,841, filed on Jun. 12, 2006, entitled “APPARATUS AND METHOD FORTHERAPEUTICALLY TREATING HUMAN AND ANIMAL CELLS, TISSUES, ORGANS ANDMOLECULES WITH ELECTROMAGNETIC FIELDS FOR TREATMENT OF DISEASES OF THEEYE AND PROPHYLACTIC TREATMENT OF THE EYE.”

U.S. patent application Ser. No. 12/819,956 is also acontinuation-in-part of U.S. patent application Ser. No. 11/903,294,filed on Sep. 20, 2007, entitled “ELECTROMAGNETIC APPARATUS FORRESPIRATORY DISEASE AND METHOD FOR USING SAME,” Publication No.US-2008-0132971-A1, now abandoned, which claims the benefit under 35U.S.C. §119 of U.S. Provisional Patent Application No. 60/846,126, filedon Sep. 20, 2006, entitled “APPARATUS AND METHOD FOR THE TREATMENT OFDISEASES OF THE LUNGS WITH ELECTROMAGNETIC FIELDS.”

U.S. patent application Ser. No. 12/819,956 is also acontinuation-in-part of U.S. patent application Ser. No. 11/977,043,filed on Oct. 22, 2007, entitled “APPARATUS AND METHOD FOR THE TREATMENTOF EXCESSIVE FIBROUS CAPSULE FORMATION AND CAPSULAR CONTRACTURE WITHELECTROMAGNETIC FIELDS,” Publication No. US-2008-0140155-A1, nowabandoned, which claims the benefit under 35 U.S.C. §119 of U.S.Provisional Patent Application No. 60/852,927, filed on Oct. 20, 2006,entitled “APPARATUS AND METHOD FOR THE TREATMENT OF EXCESSIVE FIBROUSCAPSULE FORMATION AND CAPSULAR CONTRACTURE WITH ELECTROMAGNETIC FIELDS.”

This application is related to U.S. patent application Ser. No.14/171,613, filed on Feb. 3, 2014, entitled “APPARATUS AND METHOD FORELECTROMAGNETIC TREATMENT OF NEURODEGENERATIVE CONDITIONS,” PublicationNo. US-2014-0221726-A1 and U.S. patent application Ser. No. 14/171,644,filed on Feb. 3, 2014, entitled “APPARATUS AND METHOD FORELECTROMAGNETIC TREATMENT OF NEUROLOGICAL PAIN,” Publication No.US-2014-0213844-A1.

Each of the above applications are herein incorporated by reference inits entirety.

INCORPORATION BY REFERENCE

All publications and patent applications mentioned in this specificationare herein incorporated by reference to the same extent as if eachindividual publication or patent application was specifically andindividually indicated to be incorporated by reference.

BACKGROUND Field of the Invention

This invention pertains generally to an apparatus and a method for invitro and in vivo therapeutic and prophylactic treatment of plant,animal, and human tissue, organs, cells and molecules. In particular, anembodiment according to the present invention pertains to use ofnon-thermal time-varying magnetic fields configured for optimal couplingto target pathway structures such as molecules, cells, tissue, andorgans, using power and amplitude comparison analysis to evaluate asignal to thermal noise ratio (“SNR”) in the target pathway structure.Another embodiment according to the present invention pertains toapplication of bursts of arbitrary waveform electromagnetic signals totarget pathway structures such as molecules, cells, tissues, and organsusing ultra lightweight portable coupling devices such as inductors andelectrodes, and driver circuitry that can be incorporated into apositioning device such as knee, elbow, lower back, shoulder, foot, andother anatomical wraps, as well as apparel such as garments, footware,and fashion accessories.

Yet another embodiment according to the present invention pertains toapplication of steady state periodic signals of arbitrary waveformelectromagnetic signals to target pathway structures such as molecules,cells, tissues, and organs. Examples of therapeutic and prophylacticapplications of the present invention are musculoskeletal pain relief,edema reduction, increased local blood flow, microvascular bloodperfusion, wound repair, bone repair, osteoporosis treatment andprevention, angiogenesis, neovascularization, enhanced immune response,tissue repair, enhanced transudation, and enhanced effectiveness ofpharmacological agents. An embodiment according to the present inventioncan also be used in conjunction with other therapeutic and prophylacticprocedures and modalities such as heat, cold, ultrasound, vacuumassisted wound closure, wound dressing, orthopedic fixation devices, andsurgical interventions.

This invention may also pertain generally to an electromagnetictreatment induction apparatus and a method for using same to achievemodification of cellular and tissue growth, repair, maintenance, andgeneral behavior by application of encoded electromagnetic information.More particularly this invention relates to the application ofsurgically non-invasive coupling of highly specific electromagneticsignal patterns to any number of body parts. In particular, anembodiment according to the present invention pertains to using aninduction means such as a coil to deliver pulsing electromagnetic fields(“PEMF”) to enhance living tissue growth and repair in conjunction withdevices such as supports, wraps, beds, and wheelchairs, and inconjunction with other therapeutic and wellness physical modalities,such as ultrasound, negative or positive pressure, heat, cold, massage.

This invention may also pertain generally to an apparatus and a methodfor treatment of living tissues and cells by altering their interactionwith their electromagnetic environment. This invention also relates to amethod of modification of cellular and tissue growth, repair,maintenance, and general behavior by application of encodedelectromagnetic information. More particularly this invention relates tothe application of surgically non-invasive coupling of highly specificelectromagnetic signal patterns to any number of body parts. Inparticular, an embodiment according to the present invention pertains tousing pulsing electromagnetic fields (“PEMF”) to enhance living tissuegrowth and repair via angiogenesis and neovascularization by affectingthe precursors to growth factors and other cytokines, such as ion/ligandbinding such as calcium binding to calmodoulin.

This invention may also generally relate to augmenting wound repair inhumans, plants, and animals by altering the interaction with theelectromagnetic environment of living tissues, cells, and molecules. Theinvention also relates to a method of modification of cellular andtissue growth, repair, maintenance and general behavior by theapplication of encoded electromagnetic information. More particularly,this invention provides for an application of highly specificelectromagnetic frequency (“EMF”) signal patterns to one or more bodyparts by surgically non-invasive reactive coupling of encodedelectromagnetic information. Such application of electromagneticwaveforms to human, animal, and plant target pathway structures such ascells, organs, tissues and molecules, can serve to enhance wound repair.

The use of most low frequency EMF has been in conjunction withapplications of bone repair and healing. As such, EMF waveforms andcurrent orthopedic clinical use of EMF waveforms comprise relatively lowfrequency components and are of a very low power, inducing maximumelectrical fields in a millivolts per centimeter (mV/cm) range atfrequencies under five KHz. A linear physicochemical approach employingan electrochemical model of cell membranes to predict a range of EMFwaveform patterns for which bioeffects might be expected is based uponan assumption that cell membranes, and specifically ion binding atstructures in or on cell membranes, are a likely EMF target. Therefore,it is necessary to determine a range of waveform parameters for which aninduced electric field could couple electrochemically at a cellularsurface, such as by employing voltage-dependent kinetics. Extension ofthis linear model involves Lorentz force considerations that eventuallydemonstrated that the magnetic component of EMF could play a significantrole in EMF therapeutics. This led to the ion cyclotron resonance andquantum models that predicts benefits from combined AC and DC magneticfield effects at very low frequency ranges.

The within invention is based upon biophysical and animal studies thatattribute effectiveness of cell-to-cell communication on tissuestructures' sensitivity to induced voltages and associated currents. Amathematical analysis using at least one of a Signal to Noise Ratio(“SNR”) and a Power Signal to Noise Ratio (“Power SNR”) evaluateswhether EMF signals applied to target pathway structures such as cells,tissues, organs, and molecules, are detectable above thermal noisepresent at an ion binding location. Prior art of EMF dosimetry did nottaken into account dielectric properties of tissue structures, ratherthe prior art utilized properties of isolated cells. By utilizingdielectric properties, reactive coupling of electromagnetic waveformsconfigured by optimizing SNR and Power SNR mathematical values evaluatedat a target pathway structure can enhance repair of various wounds inhuman, animal and plant cells, organs, tissues and molecules for examplepost-surgical and traumatic wound repair, angiogenesis, improved bloodperfusion, vasodilation, vasoconstriction, edema reduction, enhancedneovascularization, bone repair, tendon repair, ligament repair, organregeneration and pain relief. Wound repair enhancement results fromincreased blood flow and modulation of angiogenesis andneovascularization as well as from other enhanced bioeffectiveprocesses.

Recent clinical use of non-invasive PRF at radio frequencies has usedpulsed bursts of a 27.12 MHz sinusoidal wave, each pulse burst typicallyexhibiting a width of sixty five microseconds and having approximately1,700 sinusoidal cycles per burst, and with various burst repetitionrates.

Broad spectral density bursts of electromagnetic waveforms having afrequency in the range of one to one hundred megahertz (MHz), with 1 to100,000 pulses per burst, and with a burst-repetition rate of 0.01 to10,000 Hertz (Hz), are selectively applied to human, animal and plantcells, organs, tissues and molecules. The voltage-amplitude envelope ofeach pulse burst is a function of a random, irregular, or other likevariable, effective to provide a broad spectral density within the burstenvelope. The variables are defined by mathematical functions that takeinto account signal to thermal noise ratio and Power SNR in specifictarget pathway structures. The waveforms are designed to modulate livingcell growth, condition and repair. Particular applications of thesesignals include, but are not limited to, enhancing treatment of organs,muscles, joints, skin and hair, post surgical and traumatic woundrepair, angiogenesis, improved blood perfusion, vasodilation,vasoconstriction, edema reduction, enhanced neovascularization, bonerepair, tendon repair, ligament repair, organ regeneration and painrelief. The application of the within electromagnetic waveforms canserve to enhance healing of various wounds.

According to an embodiment of the present invention a pulse burstenvelope of higher spectral density can more efficiently couple tophysiologically relevant dielectric pathways, such as cellular membranereceptors, ion binding to cellular enzymes, and general transmembranepotential changes. An embodiment according to the present inventionincreases the number of frequency components transmitted to relevantcellular pathways, resulting in a larger range of biophysical phenomenaapplicable to known healing mechanisms becoming accessible, includingenhanced enzyme activity, growth factor release and cytokine release. Byincreasing burst duration and by applying a random, or other highspectral density envelope, to a pulse burst envelope of mono- orbi-polar rectangular or sinusoidal pulses that induce peak electricfields between 10⁻⁶ and 10 volts per centimeter (V/cm), and that satisfydetectability requirements according to SNR or Power SNR, a moreefficient and greater effect could be achieved on biological healingprocesses applicable to both soft and hard tissues in humans, animalsand plants resulting in an acceleration of wound repair.

The present invention relates to known mechanisms of wound repair thatinvolve the naturally timed release of the appropriate growth factor orcytokine in each stage of wound repair as applied to humans, animals andplants. Specifically, wound repair involves an inflammatory phase,angiogenesis, cell proliferation, collagen production, and remodelingstages. There are timed releases of specific cytokines and growthfactors in each stage. Electromagnetic fields can enhance blood flow andenhance the binding of ions which, in turn, can accelerate each healingphase. It is the specific intent of this invention to provide animproved means to enhance the action of exogenous factors and acceleraterepair. An advantageous result of using the present invention is thatwound repair can be accelerated due to enhanced blood flow or enhancedbiochemical activity. It is an object of the present invention toprovide an improved means to accelerate the intended effects or improveefficacy as well as other effects of the cytokines and growth factorsrelevant to each stage of wound repair.

Another object of the present invention is to cause and acceleratehealing of chronic wounds such as diabetic ulcers, venous stasis ulcers,pressure sores and non-healing wounds of any origin.

Another object of the present invention is that by applying a highspectral density voltage envelope as a modulating or pulse-burstdefining parameter according to SNR and Power SNR requirements, powerrequirements for such increased duration pulse bursts can besignificantly lower than that of shorter pulse bursts having pulseswithin the same frequency range; this results from more efficientmatching of frequency components to a relevant cellular/molecularprocess. Accordingly, the advantages, of enhanced transmitted dosimetryto relevant dielectric pathways and of decreased power requirements areachieved.

Therefore, a need exists for an apparatus and a method that moreeffectively accelerates wound repair in human, animal and plant cells,organs, tissues and molecules.

This invention may also relate to enhancing effectiveness ofpharmacological, chemical, cosmetic and topical agents used to treatliving tissues, cells and molecules by altering the interaction with theelectromagnetic environment of the living tissues, cells, and molecules.The invention also relates to a method of modification of cellular andtissue growth, repair, maintenance and general behavior by theapplication of encoded electromagnetic information. More particularly,this invention provides for an application of highly specificelectromagnetic frequency (“EMF”) signal patterns to one or more bodyparts by surgically non-invasive reactive coupling of encodedelectromagnetic information. Such application of electromagneticwaveforms in conjunction with pharmacological, chemical, cosmetic andtopical agents as applied to, upon, or in human, animal, and planttarget pathway structures such as cells, organs, tissues and molecules,can serve to enhance various effects of such agents.

By utilizing dielectric properties, reactive coupling of electromagneticwaveforms configured by optimizing SNR and Power SNR mathematical valuesevaluated at a target pathway structure can enhance various effects ofpharmacological, chemical, cosmetic and topical agents that are appliedto, upon or in human, animal and plant cells, organs, tissues andmolecules. An enhancement results from increased blood flow andmodulation of angiogenesis and neovascularization as well as from otherenhanced bioeffective processes.

Particular applications of these signals include, but are not limitedto, enhancing the effects of pharmacological, chemical, cosmetic andtopical agents, prophylactic and wellness treatment of organs, muscles,joints, skin and hair, post surgical and traumatic wound repair,angiogenesis, improved blood perfusion, vasodilation, vasoconstriction,edema reduction, enhanced neovascularization, bone repair, tendonrepair, ligament repair, organ regeneration and pain relief. Theapplication of the within electromagnetic waveforms in conjunction withpharmacological, chemical, cosmetic and topical agents as applied to,upon or in human, animal and plant cells, organs, tissues and moleculescan serve to enhance various effects of such compounds.

By increasing burst duration and by applying a random, or other highspectral density envelope, to a pulse burst envelope of mono- orbi-polar rectangular or sinusoidal pulses that induce peak electricfields between 10⁻⁶ and 10 volts percentimeter (V/cm), and that satisfydetectability requirements according to SNR or Power SNR, a moreefficient and greater effect could be achieved on biological healingprocesses applicable to both soft and hard tissues in humans, animalsand plants resulting in enhancement of the effectiveness ofpharmacological, chemical, cosmetic, and topical agents.

The present invention relates to known mechanisms of pharmacological,chemical, cosmetic and topical agents as applied to, upon or in human,animal and plant cells, organs, tissues and molecules. Specifically, theagents' efficacy depends upon arrival of optimal dosages of the agentsto intended target pathway structures, which can be accomplished eithervia enhanced blood flow or enhanced chemical activity catalyzed by anincrease in active enzymes during a relevant biochemical cascade.Electromagnetic fields can enhance blood flow and ion binding whichaffect the agents' activity. An advantageous result of using the presentinvention is that the quantity of an agent may be able to be reduced dueto the agents enhanced effectiveness. It is an object of the presentinvention to provide an improved means to enhance and accelerate theintended effects, and improve efficacy as well as other effects ofpharmacological, chemical, cosmetic and topical agents applied to, uponor in human, animal and plant cells, organs, tissues and molecules.

Therefore, a need exists for an apparatus and a method that moreeffectively enhances and accelerates the intended effects, and improveefficacy as well as other bioeffective effects of pharmacological,chemical, cosmetic and topical agents applied to, upon or in human,animal and plant cells, organs, tissues and molecules.

This invention may also pertain generally to an electromagnetictreatment integrated coil apparatus and a method for using same toachieve modification of cellular and tissue growth, repair, maintenance,and general behavior by application of encoded electromagneticinformation. More particularly this invention relates to the applicationof surgically non-invasive coupling of highly specific electromagneticsignal patterns to any number of body parts. This invention also relatesto treatment of living tissues and cells by altering their interactionwith their electromagnetic environment. The invention further relates toa method of modification of cellular and tissue growth, repair,maintenance, and general behavior by the application of encodedelectromagnetic information. In particular, an embodiment according tothe present invention pertains to using an induction means such as acoil to deliver pulsing electromagnetic fields (“PEMF”) to enhanceliving tissue growth and repair integrated with devices such assupports, wraps, beds, and wheelchairs, and in conjunction with othertherapeutic and wellness physical modalities, such as ultrasound,negative or positive pressure, heat, cold, massage.

This invention may also pertain generally to an apparatus and a methodfor using electromagnetic therapy treatment for hair maintenance andrestoration and for treatment of degenerative neurological pathologiesand other cerebrofacial conditions, including sleep disorders, bymodulation of the interaction of hair, cerebral, neurological, and othertissues with their in situ electromagnetic environment. This inventionalso relates to a method of modification of cellular and tissue growth,repair, maintenance, and general behavior by application of encodedelectromagnetic information to molecules, cells, tissues and organs onhumans and animals. More particularly this invention relates to theapplication of surgically non-invasive coupling of highly specificelectromagnetic signal patterns to hair and other cerebrofacial tissue.In particular, an embodiment according to the present invention pertainsto using a self-contained apparatus that emits time varying magneticfields (“PMF”) configured using specific mathematical models to enhancehair and other tissue growth and repair by affecting the initial stepsto growth factors and other cytokine release, such as ion/ligand bindingfor example calcium binding to calmodoulin.

This invention may relates to delivering electromagnetic signals toophthalmic tissue of humans and animals that are injured or diseasedwhereby the interaction with the electromagnetic environment of livingtissues, cells, and molecules is altered to achieve a therapeutic orwellness effect. The invention also relates to a method of modificationof cellular and tissue growth, repair, maintenance and general behaviorby the application of encoded electromagnetic information. Moreparticularly, this invention provides for an application of highlyspecific electromagnetic frequency (“EMF”) signal patterns to ophthalmictissue by surgically non-invasive reactive coupling of encodedelectromagnetic information. Such application of electromagneticwaveforms to human and animal target pathway structures such as cells,organs, tissues and molecules, can serve to remedy injured or diseasedophthalmic tissue or to prophylactically treat such tissue.

The use of most low frequency EMF has been in conjunction withapplications of bone repair and healing. As such, EMF waveforms andcurrent orthopedic clinical use of EMF waveforms comprise relatively lowfrequency components inducing maximum electrical fields in a millivoltsper centimeter (mV/cm) range at frequencies under five KHz. A linearphysicochemical approach employing an electrochemical model of cellmembranes to predict a range of EMF waveform patterns for whichbioeffects might be expected is based upon an assumption that cellmembranes, and specifically ion binding at structures in or on cellmembranes or surfaces, are a likely EMF target. Therefore, it isnecessary to determine a range of waveform parameters for which aninduced electric field could couple electrochemically at a cellularsurface, such as by employing voltage-dependent kinetics.

The within invention is based upon biophysical and animal studies thatattribute effectiveness of cell-to-cell communication on tissuestructures' sensitivity to induced voltages and associated currents. Amathematical analysis using at least one of a Signal to Noise Ratio(“SNR”) and a Power Signal to Noise Ratio (“Power SNR”) evaluateswhether EMF signals applied to target pathway structures such as cells,tissues, organs, and molecules, are detectable above thermal noisepresent at an ion binding location. Prior art of EMF dosimetry did nottake into account dielectric properties of tissue structures, rather theprior art utilized properties of isolated cells. By utilizing dielectricproperties, reactive coupling of electromagnetic waveforms configured byoptimizing SNR and Power SNR mathematical values evaluated at a targetpathway structure can enhance wellness of the ophthalmic system as wellas repair of various ophthalmic injuries and diseases in human andanimal cells, organs, tissues and molecules for example wet maculardegeneration and dry macular degeneration. Cell, organ, tissue, andmolecule repair enhancement results from increased blood flow andanti-inflammatory effects, and modulation of angiogenesis andneovascularization as well as from other enhanced bioeffective processessuch as growth factor and cytokine release.

Broad spectral density bursts of electromagnetic waveforms having afrequency in the range of one hertz (Hz) to one hundred megahertz (MHz),with 1 to 100,000 pulses per burst, and with a burst-repetition rate of0.01 to 10,000 Hertz (Hz), are selectively applied to human and animalcells, organs, tissues and molecules. The voltage-amplitude envelope ofeach pulse burst is a function of a random, irregular, or other likevariable, effective to provide a broad spectral density within the burstenvelope. The variables are defined by mathematical functions that takeinto account signal to thermal noise ratio and Power SNR in specifictarget pathway structures. The waveforms are designed to modulate livingcell growth, condition and repair. Particular applications of thesesignals include, but are not limited to, enhancing treatment of organs,muscles, joints, eyes, skin and hair, post surgical and traumatic woundrepair, angiogenesis, improved blood perfusion, vasodilation,vasoconstriction, edema reduction, enhanced neovascularization, bonerepair, tendon repair, ligament repair, organ regeneration and painrelief. The application of the within electromagnetic waveforms canserve to enhance healing of various ophthalmic tissue injuries anddiseases, as well as provide prophylactic treatment for such tissue.

According to an embodiment of the present invention a pulse burstenvelope of higher spectral density can more efficiently couple tophysiologically relevant dielectric pathways, such as cellular membranereceptors, ion binding to cellular enzymes, and general transmembranepotential changes. An embodiment according to the present inventionincreases the number of frequency components transmitted to relevantcellular pathways, resulting in different electromagneticcharacteristics of healing tissue and a larger range of biophysicalphenomena applicable to known healing mechanisms becoming accessible,including enhanced enzyme activity, second messenger, such as nitricoxide (“NO”) release, growth factor release and cytokine release. Byincreasing burst duration and by applying a random, or other highspectral density envelope, to a pulse burst envelope of mono-polar orbi-polar rectangular or sinusoidal pulses that induce peak electricfields between 10⁻⁶ and 10 volts per centimeter (V/cm), and that satisfydetectability requirements according to SNR or Power SNR, a moreefficient and greater effect could be achieved on biological healingprocesses applicable to both soft and hard tissues in humans and animalsresulting in an acceleration of ophthalmic injury and disease repair.

The present invention relates to known mechanisms of ophthalmic injuryand disease repair and healing that involve the naturally timed releaseof the appropriate anti-inflammatory cascade and growth factor orcytokine release in each stage of wound repair as applied to humans andanimals. Specifically, ophthalmic injury and disease repair involves aninflammatory phase, angiogenesis, cell proliferation, collagenproduction, and remodeling stages. There are timed releases of secondmessengers, such as NO, specific cytokines and growth factors in eachstage. Electromagnetic fields can enhance blood flow and enhance thebinding of ions, which, in turn, can accelerate each healing phase. Itis the specific intent of this invention to provide an improved means toenhance the action of endogenous factors and accelerate repair and toaffect wellness. An advantageous result of using the present inventionis that ophthalmic injury and disease repair, and healing can beaccelerated due to enhanced blood flow or enhanced biochemical activity.In particular, an embodiment according to the present invention pertainsto using an induction means such as a coil to deliver pulsingelectromagnetic fields (“PEMF”) for the maintenance of the ophthalmicsystem and the treatment of ophthalmic diseases such as maculardegeneration, glaucoma, retinosa pigmentosa, repair and regeneration ofoptic nerve prophylaxis, and other related diseases. More particularly,this invention provides for the application, by surgically non-invasivereactive coupling, of highly specific electromagnetic signal patterns toone or more body parts. Such applications made on a non-invasive basisto the constituent tissues of the ophthalmic system and its surroundingtissues can serve to improve the physiological parameters of ophthalmicdiseases.

An object of the present invention may be to provide an improved meansto accelerate the intended effects or improve efficacy as well as othereffects of the second messengers, cytokines and growth factors relevantto each stage of ophthalmic injury and disease repair and healing.

Another object of the present invention may be to cause and acceleratehealing for treatment of ophthalmic diseases such as wet maculardegeneration, dry macular degeneration, glaucoma, retinosa pigmentosa,repair and regeneration of optic nerve, prophylaxis, and other relateddiseases.

Another object of the present invention may be to accelerate healing ofophthalmic injuries of any type.

Another object of the present invention is to maintain wellness of theophthalmic system.

Another object of the present invention is that by applying a highspectral density voltage envelope as a modulating or pulse-burstdefining parameter according to SNR and Power SNR requirements, powerrequirements for such increased duration pulse bursts can besignificantly lower than that of shorter pulse bursts having pulseswithin the same frequency range; this results from more efficientmatching of frequency components to a relevant cellular/molecularprocess. Accordingly, the advantages of enhanced transmitted dosimetryto relevant dielectric pathways and of decreased power requirements, areachieved.

Therefore, a need exists for an apparatus and a method that effectivelyenhances wellness of the ophthalmic system and accelerates healing ofophthalmic injuries, ophthalmic diseases, areas around the ophthalmicsystem by modulating ion binding at cells, organs, tissues and moleculesof humans and animals.

This invention may pertain to delivering electromagnetic signals torespiratory tissue such as lung tissue, of humans and animals that areinjured or diseased whereby the interaction with the electromagneticenvironment of living tissues, cells, and molecules is altered toachieve a therapeutic or wellness effect. The invention also relates toa method of modification of cellular and tissue growth, repair,maintenance and general behavior by the application of encodedelectromagnetic information. More particularly, this invention providesfor an application of highly specific electromagnetic frequency (“EMF”)signal patterns to lung tissue by surgically non-invasive reactivecoupling of encoded electromagnetic information. Such application ofelectromagnetic waveforms to human and animal target pathway structuressuch as cells, organs, tissues and molecules, can serve to remedyinjured or diseased respiratory tissue or to prophylactically treat suchtissue.

The use of most low frequency EMF has been in conjunction withapplications of bone repair and healing. As such, EMF waveforms andcurrent orthopedic clinical use of EMF waveforms comprise relatively lowfrequency components inducing maximum electrical fields in a millivoltsper centimeter (mV/cm) range at frequencies under five KHz. A linearphysicochemical approach employing an electrochemical model of cellmembranes to predict a range of EMF waveform patterns for whichbioeffects might be expected is based upon an assumption that cellmembranes, and specifically ion binding at structures in or on cellmembranes or surfaces, are a likely EMF target. Therefore, it isnecessary to determine a range of waveform parameters for which aninduced electric field could couple electrochemically at a cellularsurface, such as by employing voltage-dependent kinetics.

A pulsed radio frequency (“PRF”) signal derived from a 27.12 MHzcontinuous sine wave used for deep tissue healing is known in the priorart of diathermy. A pulsed successor of the diathermy signal wasoriginally reported as an electromagnetic field capable of eliciting anon-thermal biological effect in the treatment of infections.Subsequently, PRF therapeutic applications have been reported for thereduction of post-traumatic and post-operative pain and edema in softtissues, wound healing, burn treatment, and nerve regeneration. Theapplication of PRF for resolution of traumatic and chronic edema hasbecome increasingly used in recent years. Results to date using PRF inanimal and clinical studies suggest that edema may be measurably reducedfrom such electromagnetic stimulus

The within inventions may be based upon biophysical and animal studiesthat attribute effectiveness of cell-to-cell communication on tissuestructures' sensitivity to induced voltages and associated currents. Amathematical power comparison analysis using at least one of a Signal toNoise Ratio (“SNR”) and a Power Signal to Noise Ratio (“Power SNR”)evaluates whether EMF signals applied to target pathway structures suchas cells, tissues, organs, and molecules, are detectable above thermalnoise present at an ion binding location. Prior art of EMF dosimetry didnot take into account dielectric properties of tissue structures, ratherthe prior art utilized properties of isolated cells. By utilizingdielectric properties, reactive coupling of electromagnetic waveformsconfigured by optimizing SNR and Power SNR mathematical values evaluatedat a target pathway structure can enhance wellness of the respiratorysystem as well as repair of various respiratory injuries and diseases inhuman and animal cells, organs, tissues and molecules for examplesarcoidosis, granulomatous pneumonitis, pulmonary fibrosis, and “WorldTrade Center Cough.” Cell, organ, tissue, and molecule repairenhancement results from increased blood flow and anti-inflammatoryeffects, and modulation of angiogenesis and neovascularization as wellas from other enhanced bioeffective processes such as growth factor andcytokine release.

As mentioned above, broad spectral density bursts of electromagneticwaveforms having a frequency in the range of one hertz (Hz) to onehundred megahertz (MHz), with 1 to 100,000 pulses per burst, and with aburst-repetition rate of 0.01 to 10,000 Hertz (Hz), are selectivelyapplied to human and animal cells, organs, tissues and molecules. Thevoltage-amplitude envelope of each pulse burst is a function of arandom, irregular, or other like variable, effective to provide a broadspectral density within the burst envelope. The variables are defined bymathematical functions that take into account signal to thermal noiseratio and Power SNR in specific target pathway structures. The waveformsare designed to modulate living cell growth, condition and repair.Particular applications of these signals include, but are not limitedto, enhancing treatment of organs, muscles, joints, eyes, skin and hair,post surgical and traumatic wound repair, angiogenesis, improved bloodperfusion, vasodilation, vasoconstriction, edema reduction, enhancedneovascularization, bone repair, tendon repair, ligament repair, organregeneration and pain relief. The application of the withinelectromagnetic waveforms can serve to enhance healing of variousrespiratory tissue injuries and diseases, as well as provideprophylactic treatment for such tissue. The present invention is anon-invasive, non-pharmacological treatment modality that can have asalutary impact on persons suffering from respiratory diseases orconditions or that can be used on a prophylactic basis for thoseindividuals who may be prone to respiratory diseases or conditions.

An aspect of the present invention is that a pulse burst envelope ofhigher spectral density can more efficiently couple to physiologicallyrelevant dielectric pathways, such as cellular membrane receptors, ionbinding to cellular enzymes, and general transmembrane potentialchanges. Another aspect of the present invention increases the number offrequency components transmitted to relevant cellular pathways,resulting in different electromagnetic characteristics of healing tissueand a larger range of biophysical phenomena applicable to known healingmechanisms becoming accessible, including enhanced enzyme activity,second messenger, such as nitric oxide (“NO”) release, growth factorrelease and cytokine release. By increasing burst duration and byapplying a random, or other high spectral density envelope, to a pulseburst envelope of mono-polar or bi-polar rectangular or sinusoidalpulses that induce peak electric fields between 10⁻⁶ and 10 volts percentimeter (V/cm), and that satisfy detectability requirements accordingto SNR or Power SNR, a more efficient and greater effect could beachieved on biological healing processes applicable to both soft andhard tissues in humans and animals resulting in an acceleration ofrespiratory injury and disease repair.

The present invention relates to known mechanisms of respiratory injuryand disease repair and healing that involve the naturally timed releaseof the appropriate anti-inflammatory cascade and growth factor orcytokine release in each stage of wound repair as applied to humans andanimals. Specifically, respiratory injury and disease repair involves aninflammatory phase, angiogenesis, cell proliferation, collagenproduction, and remodeling stages. There are timed releases of secondmessengers, such as NO, specific cytokines and growth factors in eachstage. Electromagnetic fields can enhance blood flow and enhance thebinding of ions, which, in turn, can accelerate each healing phase. Itis the specific intent of this invention to provide an improved means toenhance the action of endogenous factors and accelerate repair and toaffect wellness. An advantageous result of using the present inventionis that respiratory injury and disease repair, and healing can beaccelerated due to enhanced blood flow or enhanced biochemical activity.In particular, an embodiment according to the present invention pertainsto using an induction means such as a coil to deliver pulsingelectromagnetic fields (“PEMF”) for the maintenance of the respiratorysystem and the treatment of respiratory diseases such sarcoidosis,granulomatous pneumonitis, pulmonary fibrosis, and “World Trade CenterCough,” and other related diseases. More particularly, this inventionprovides for the application, by surgically non-invasive reactivecoupling, of highly specific electromagnetic signal patterns to one ormore body parts. Such applications made on a non-invasive basis to theconstituent tissues of the respiratory system and its surroundingtissues can serve to improve the physiological parameters of respiratorydiseases.

Sarcoidosis, granulomatous pneumonitis, pulmonary fibrosis, and otherrelated diseases result from inflammatory processes caused by inhalationof foreign material into lung tissue. The initiation of such diseases isthe inflammation that occurs after particle inhalation. The withininvention produces a physiological effect designed to reduce theinflammatory response, which in turn, may reduce the effects of inhaledforeign bodies on lung capacity and even prevent other systemic healthproblems. A number of physiological cascades that are accelerated ormodified by the waveforms produced by the methods and apparatus of thisinvention serve to reduce the inflammatory processes. In particular, thePEMF signal can enhance the production of nitric oxide via modulation ofCalcium (“Ca²⁺”) binding to calmodulin (“CaM”). This in turn can inhibitinflammatory leukotrienes that reduce the inflammatory process leadingto excessive fibrous tissue for example scars, in lung tissue.Prophylactic use of the within invention by first responders may preventor reduce the inflammatory processes leading to formation of fibroustissue leading to lung disease.

Sarcoidosis involves inflammation that produces tiny agglomerations ofcells in various organs of the body. These agglomerations are calledglanulomas which are an aggregation and proliferation of macrophages toform nodules or granules. Such granulomas are of microscopic size andare not easily identifiable without significant magnification.Granulomas can grow and join together creating large and small groups ofagglomerated cells. If there is a high prevalence of agglomeratedgranulomas in an organ, such as the lungs, the agglomerated granulomascan negatively impact the proper functioning of that organ. In thelungs, this negative impact can cause symptoms of sarcoidosis.Sarcoidosis can occur in almost any part of the body although it usuallyaffects some organs such as the lungs and lymphnodes, more than others.It usually begins in one or two places, the lungs or lymphnodesespecially the lymphnodes in the chest cavity. Sarcoidosis almost alwaysoccurs in more than one organ at a time. Exposure to pollutants or otherparticulates that are breathed into the lungs, such as dust and fiberspresent at the World Trade Center site after Sep. 11, 2001, can causethe scarring and resultant sarcoidosis.

Sarcoidosis involves both an active and a non-active phase. In theactive phase, granulomas are formed and grow with symptoms developing.Scar tissue can form in the organs where such granulomas occur andinflammation is present. In the non-active phase, inflammation reduces,and the granulomas do not grow or may be reduced in size. If thenon-active phase does occur, any scarring that occurred will remain andcause increased or continuing symptoms.

The course of the disease varies greatly. Sarcoidosis may be mild orsevere. The inflammation that causes the granulomas may resolve withoutintervention and may stop growing or reduce in size. Symptoms may bereduced or alleviated within a few years after onset. In some cases, theinflammation remains but does not progress. There may be increasedsymptoms or flare-ups that require treatment on an intermittent basis.Although drug intervention can help, sarcoidosis may leave scar tissuein the lungs, skin, eyes or other organs and that scar tissue canpermanently affect the functioning of the organs. Drug treatment usuallydoes not affect scar tissue. The present invention has been shown inanimal and clinical testing to reduce inflammation and accelerateangiogenesis and revascularization in organ tissue that may lead toimprovement of vascularity of the tissue surrounding the scarring thatmay be the result of sarcoidosis in the lungs.

Sarcoidosis usually occurs slowly over many months and does not usuallycause sudden illness. However, some symptoms may occur suddenly. Thesesymptoms include disturbed heart rhythms, arthritis in the ankles, andeye symptoms. In some serious cases in which vital organs are affected,sarcoidosis can resulting death. However, sarcoidosis is not a form ofcancer. Presently there is no way to prevent sarcoidosis. Sarcoidosiswas once thought to be an uncommon condition. It is now known to affecttens of thousands of people throughout the United States. Since manypeople who have sarcoidosis exhibit no symptoms, it is difficult todetermine the actual prevalence of sarcoidosis in populations, althoughthere seems to be a higher incidence in certain cultures.

An aspect of the present invention is to provide an improved means toaccelerate the intended effects or improve efficacy as well as othereffects of the second messengers, cytokines and growth factors relevantto each stage of respiratory injury and disease repair and healing.

Another aspect of the present invention is to cause and acceleratehealing for treatment of respiratory diseases such as, sarcoidosis,granulomatous pneumonitis, pulmonary fibrosis, and “World Trade CenterCough” and other related diseases.

Another aspect of the present invention is to accelerate healing ofrespiratory injuries of any type.

Another aspect of the present invention is to maintain wellness of therespiratory system.

Another aspect of the present invention is that by applying a highspectral density voltage envelope as a modulating or pulse-burstdefining parameter according to SNR and Power SNR requirements, powerrequirements for such increased duration pulse bursts can besignificantly lower than that of shorter pulse bursts having pulseswithin the same frequency range; this results from more efficientmatching of frequency components to a relevant cellular/molecularprocess. Accordingly, the advantage of enhanced transmitted dosimetry torelevant dielectric pathways and the advantage of decreased powerrequirements, are achieved. This advantageously allows forimplementation of the within invention in an easily transportable unitfor ease of application to the lung area and is particularly suitablefor prophylactic use by first responders.

Another aspect of the present invention allows application of specificwaveforms in a convenient and comfortable configuration to a desiredpulmonary area. In an embodiment according to the present invention, aportable generator with multiple coil applicators that are incorporatedinto a body-conforming garment is worn by the user during a posterioritreatment or worn prophylactically. This allows for the properpositioning of the output coils to the chest area thereby allowing theproduced signals to be broadcast over the lungs in an efficient manner.

Therefore, a need exists for an apparatus and a method that effectivelyenhances wellness of the respiratory system and accelerates healing ofrespiratory injuries, respiratory diseases, and areas around therespiratory system by modulating ion binding at cells, organs, tissuesand molecules of humans and animals.

This invention pertains generally to an electromagnetic coil apparatusand a method that configures and delivers electromagnetic signals topromote cell and tissue growth, repair, and maintenance. Theelectromagnetic environment of living tissues, cells, and molecules isaltered by the electromagnetic signals generated by an embodiment of thepresent invention to achieve a therapeutic or wellness effect.Alteration of the electromagnetic environment can be particularlyeffective for alleviating pain and discomfort in individuals havingcapsular contracture or excessive fibrous capsule formation associatedwith any surgically implanted device. The invention also relates to amethod of modification of cellular and tissue growth, repair,maintenance and general behavior by the application of encodedelectromagnetic information. More particularly, this invention providesfor an application of highly specific electromagnetic frequency (“EMF”)signal patterns to excessive fibrous capsule tissue by non-invasivereactive coupling of encoded electromagnetic information. Suchapplication of electromagnetic waveforms to human and animal fibrouscapsule formation and capsular contracture target pathway structuressuch as cells, organs, tissues and molecules, can reduce the pain andedema associated with capsular contracture, can increase blood flow,neovascularization, vasculargenesis, and angiogenesis and can augmentthe release of growth factors and cytokines related to the prophylacticand a posteriori treatment of excessive fibrous capsule formation.

The present invention further relates to altering the cellular andmolecular mechanisms of excessive fibrous capsule formation and tocontrol capsular contracture generally associated with post surgicalcomplications of implants such as breast augmentation.

Capsular contracture is a painful inflammatory condition which can occurat any time post surgically but usually occurs within the first severalmonths after surgery. Capsular contracture is the most commoncomplication of breast augmentation surgery but also can occur withother surgically implanted devices. At the time of initial breastaugmentation surgery, a pocket is made for a breast implant in tissuecovering the chest. During the healing process a capsule that iscomprised of fibrous tissue forms. The body is genetically programmed tocounteract that formation by attempting to shrink the scar tissue to acertain degree. Under normal circumstances, the pocket remains open thusallowing the implant to look and feel natural. However in a certainnumber of cases, the capsule will tighten thereby causing pressure byrestricting the space for the implant. Furthermore this causes theimplant to feel hard and rigid with concomitant distortion of theappearance of the breast. In later stages the implant feels extremelyfirm and may take on an unnatural “ball like” appearance. The presentinvention produces a physiological effect in the tissue of a capsularcontracture. The physiological effect causes revascularization andinter-cellular modification tissue, to reduce in hardness and prevalencethereby reducing pain and discomfort for a patient. Waveforms producedby the within invention accelerate or modify a number of physiologicalcascades that either alleviate the propensity of the capsule to compressor harden, or produce a reduction in the existing capsule involvementwith the physical area at which the waveforms have been applied to. Inparticular a pulsing electromagnetic field (“PEMF”) signal can enhanceproduction of nitric oxide (“NO”) via modulation of Calcium (“Ca2+”)binding to calmodulin (“CaM”). This in turn can inhibit inflammatoryleukotrienes that reduce the inflammatory process leading to excessivefibrous capsule formation. At present, pharmacologic agents targeted toinhibit leukotrienes are employed for treating capsular contracture withlimited success. Prophylactic use of the within invention prior todevice implant in individuals that are deemed susceptible to capsularcontracture formation may prevent or reduce the formation of excessivefibrous tissue.

An advantageous result of the within invention is that by applying ahigh spectral density voltage envelope as the modulating or pulse-burstdefining parameter, the power requirement for such increased durationpulse bursts can be significantly lower than that of shorter pulsebursts containing pulses within the same frequency range. This is due tomore efficient matching of the frequency components to relevant cellularand molecular processes. Accordingly the dual advantages of enhancedtransmitted dosimetry to the relevant dielectric pathways and ofdecreased power requirements are achieved. This allows for theimplementation of the within invention in an easily transportable unitfor ease of application on capsular contracture patients.

Therefore, a need exists for an apparatus and a method that effectivelyaccelerates or modifies a number of physiological cascades thatalleviate the propensity of the capsule to compress or harden, thatreduce excessive fibrous capsule formation, and that produce a reductionin the existing capsule involvement within the physical area to whichthe waveforms have been applied.

Described herein are also electromagnetic treatment devices, systems andmethods. Some embodiments pertain generally to a method and apparatusfor therapeutic and prophylactic treatment of animal and human nervoussystem. In particular, some embodiments pertain to use of non-thermaltime-varying electromagnetic fields configured to accelerate theasymmetrical kinetics of the binding of intracellular ions to theirrespective binding proteins which regulate the biochemical signalingpathways living systems employ to contain and reduce the inflammatoryresponse to injury. Other embodiments pertain to the non-thermalapplication of repetitive pulse bursts of sinusoidal, rectangular,chaotic or arbitrary waveform electromagnetic fields to instantaneouslyaccelerate ion-buffer binding in signaling pathways in animal and humannervous system using ultra lightweight portable coupling devices such asinductors and electrodes, driven by miniature signal generator circuitrythat can be incorporated into an anatomical positioning device such as adressing, bandage, compression bandage, compression dressing; lumbar orcervical back, shoulder, head, neck and other body portion wraps andsupports; garments, hats, caps, helmets, mattress pads, seat cushions,beds, stretchers, and other body supports in cars, motorcycles, buses,trains, airplanes, boats, ships and the like.

Yet another embodiment pertains to application of sinusoidal,rectangular, chaotic or arbitrary waveform electromagnetic signals,having frequency components below about 100 GHz, configured toaccelerate the binding of intracellular Ca²⁺ to a buffer, such as CaM,to enhance biochemical signaling pathways in animal and human nervoussystem. Signals configured according to additional embodiments produce anet increase in a bound ion, such as Ca²⁺, at CaM binding sites becausethe asymmetrical kinetics of Ca/CaM binding allows such signals toaccumulate voltage induced at the ion binding site, thereby acceleratingvoltage-dependent ion binding. Examples of therapeutic and prophylacticapplications of the present invention are modulation of biochemicalsignaling in anti-inflammatory pathways, modulation of biochemicalsignaling in cytokine release pathways, modulation of biochemicalsignaling in growth factor release pathways; edema and lymph reduction,anti-inflammatory, post surgical and post operative pain and edemarelief, nerve, bone and organ pain relief, increased local blood flow,microvascular blood perfusion, treatment of tissue and organ ischemia,brain tissue ischemia from stroke or traumatic brain injury, treatmentof neurological injury and neurodegenerative diseases such asAlzheimer's and Parkinson's; angiogenesis, neovascularization; enhancedimmune response; enhanced effectiveness of pharmacological agents; nerveregeneration; prevention of apoptosis; modulation of heat shock proteinsfor prophylaxis and response to injury or pathology.

Some embodiments can also be used in conjunction with other therapeuticand prophylactic procedures and modalities such as heat, cold, light,ultrasound, mechanical manipulation, massage, physical therapy, wounddressings, orthopedic and other surgical fixation devices, and surgicalinterventions. In addition, any of the variations described herein canalso be used in conjunction with one or more pharmacological agents. Anyof the variations described herein can also be used with imaging ornon-imaging diagnostic procedures.

In some variations the systems, devices and/or methods generally relateto application of electromagnetic fields (EMF), and in particular,pulsed electromagnetic fields (PEMF), including a subset of PEMF in aradio frequency domain (e.g., pulsed radio frequency or PRF), for thetreatment of head, cerebral and neural injury, includingneurodegenerative conditions in animals and humans.

Discussion of Related Art

It is now well established that application of weak non-thermalelectromagnetic fields (“EMF”) can result in physiologically meaningfulin vivo and in vitro bioeffects. Time-varying electromagnetic fields,comprising rectangular waveforms such as pulsing electromagnetic fields(“PEMF”), and sinusoidal waveforms such as pulsed radio frequency fields(“PRF”) ranging from several Hertz to an about 15 to an about 40 MHzrange, are clinically beneficial when used as an adjunctive therapy fora variety of musculoskeletal injuries and conditions.

Beginning in the 1960's, development of modern therapeutic andprophylactic devices was stimulated by clinical problems associated withnon-union and delayed union bone fractures. Early work showed that anelectrical pathway can be a means through which bone adaptively respondsto mechanical input. Early therapeutic devices used implanted andsemi-invasive electrodes delivering direct current (“DC”) to a fracturesite. Non-invasive technologies were subsequently developed usingelectrical and electromagnetic fields. These modalities were originallycreated to provide a non-invasive “no-touch” means of inducing anelectrical/mechanical waveform at a cell/tissue level. Clinicalapplications of these technologies in orthopaedics have led to approvedapplications by regulatory bodies worldwide for treatment of fracturessuch as non-unions and fresh fractures, as well as spine fusion.Presently several EMF devices constitute the standard armamentarium oforthopaedic clinical practice for treatment of difficult to healfractures. The success rate for these devices has been very high. Thedatabase for this indication is large enough to enable its recommendeduse as a safe, non-surgical, non-invasive alternative to a first bonegraft. Additional clinical indications for these technologies have beenreported in double blind studies for treatment of avascular necrosis,tendinitis, osteoarthritis, wound repair, blood circulation and painfrom arthritis as well as other musculoskeletal injuries.

Cellular studies have addressed effects of weak low frequencyelectromagnetic fields on both signal transduction pathways and growthfactor synthesis. It can be shown that EMF stimulates secretion ofgrowth factors after a short, trigger-like duration. Ion/ligand bindingprocesses at a cell membrane are generally considered an initial EMFtarget pathway structure. The clinical relevance to treatments forexample of bone repair, is upregulation such as modulation, of growthfactor production as part of normal molecular regulation of bone repair.Cellular level studies have shown effects on calcium ion transport, cellproliferation, Insulin Growth Factor (“IGF-II”) release, and IGF-IIreceptor expression in osteoblasts. Effects on Insulin Growth Factor-I(“IGF-I”) and IGF-II have also been demonstrated in rat fracture callus.Stimulation of transforming growth factor beta (“TGF-β”) messenger RNA(“mRNA”) with PEMF in a bone induction model in a rat has been shown.Studies have also demonstrated upregulation of TGF-β mRNA by PEMF inhuman osteoblast-like cell line designated MG-63, wherein there wereincreases in TGF-β1, collagen, and osteocalcin synthesis. PEMFstimulated an increase in TGF-β1 in both hypertrophic and atrophic cellsfrom human non-union tissue. Further studies demonstrated an increase inboth TGF-β1 mRNA and protein in osteoblast cultures resulting from adirect effect of EMF on a calcium/calmodulin-dependent pathway.Cartilage cell studies have shown similar increases in TGF-β1 mRNA andprotein synthesis from EMF, demonstrating a therapeutic application tojoint repair. U.S. Pat. No. 4,315,503 (1982) to Ryaby and U.S. Pat. No.5,723,001 (1998) to Pilla typify the research conducted in this field.

However, prior art in this field applies unnecessarily high amplitudeand power to a target pathway structure, requires unnecessarily longtreatment time, and is not portable.

Therefore, a need exists for an apparatus and a method that moreeffectively modulates biochemical processes that regulate tissue growthand repair, shortens treatment times, and incorporates miniaturizedcircuitry and light weight applicators thus allowing the apparatus to beportable and if desired disposable. A further need exists for anapparatus and method that more effectively modulates biochemicalprocesses that regulate tissue growth and repair, shortens treatmenttimes, and incorporates miniaturized circuitry and light weightapplicators that can be constructed to be implantable.

EMF has been used in applications of bone repair and bone healing.Waveforms comprising low frequency components and low power arecurrently used in orthopedic clinics. Origins of using bone repairsignals began by considering that an electrical pathway may constitute ameans through which bone can adaptively respond to EMF signals. A linearphysicochemical approach employing an electrochemical model of a cellmembrane predicted a range of EMF waveform patterns for which bioeffectsmight be expected. Since a cell membrane was a likely EMF target, itbecame necessary to find a range of waveform parameters for which aninduced electric field could couple electrochemically at the cellularsurface, such as voltage-dependent kinetics. Extension of this linearmodel also involved Lorentz force analysis.

A pulsed radio frequency (“PRF”) signal derived from a 27.12 MHzcontinuous sine wave used for deep tissue healing is known in the priorart of diathermy. A pulsed successor of the diathermy signal wasoriginally reported as an electromagnetic field capable of eliciting anon-thermal biological effect in the treatment of infections. PRFtherapeutic applications have been reported for reduction ofpost-traumatic and post-operative pain and edema in soft tissues, woundhealing, burn treatment and nerve regeneration. Application of EMF forthe resolution of traumatic edema has become increasingly used in recentyears. Results to date using PRF in animal and clinical studies suggestthat edema may be measurably reduced from such electromagnetic stimulus.

Prior art considerations of EMF dosimetry have not taken into accountdielectric properties of tissue structure as opposed to the propertiesof isolated cells.

In recent years, clinical use of non-invasive PRF at radio frequenciescomprised using pulsed bursts of a 27.12 MHz sinusoidal wave, whereineach pulse burst comprises a width of sixty-five microseconds, havingapproximately 1,700 sinusoidal cycles per burst, and various burstrepetition rates. This limited frequency components that could couple torelevant dielectric pathways in cells and tissue.

Time-varying electromagnetic fields, comprising rectangular waveformssuch as pulsing electromagnetic fields, and sinusoidal waveforms such aspulsed radio frequency fields ranging from several Hertz to an about 15to an about 40 MHz range, are clinically beneficial when used as anadjunctive therapy for a variety of musculoskeletal injuries andconditions.

Beginning in the 1960's, development of modern therapeutic andprophylactic devices was stimulated by clinical problems associated withnon-union and delayed union bone fractures. Early work showed that anelectrical pathway can be a means through which bone adaptively respondsto mechanical input. Early therapeutic devices used implanted andsemi-invasive electrodes delivering direct current (“DC”) to a fracturesite. Non-invasive technologies were subsequently developed usingelectrical and electromagnetic fields. These modalities were originallycreated to provide a non-invasive “no-touch” means of inducing anelectrical/mechanical waveform at a cell/tissue level. Clinicalapplications of these technologies in orthopaedics have led to approvedapplications by regulatory bodies worldwide for treatment of fracturessuch as non-unions and fresh fractures, as well as spine fusion.Presently several EMF devices constitute the standard armamentarium oforthopaedic clinical practice for treatment of difficult to healfractures. The success rate for these devices has been very high. Thedatabase for this indication is large enough to enable its recommendeduse as a safe, non-surgical, non-invasive alternative to a first bonegraft. Additional clinical indications for these technologies have beenreported in double blind studies for treatment of avascular necrosis,tendinitis, osteoarthritis, wound repair, blood circulation and painfrom arthritis as well as other musculoskeletal injuries.

Cellular studies have addressed effects of weak low frequencyelectromagnetic fields on both signal transduction pathways and growthfactor synthesis. It can be shown that EMF stimulates secretion ofgrowth factors after a short, trigger-like duration. Ion/ligand bindingprocesses at a cell membrane are generally considered an initial EMFtarget pathway structure. The clinical relevance to treatments forexample of bone repair, is upregulation such as modulation, of growthfactor production as part of normal molecular regulation of bone repair.Cellular level studies have shown effects on calcium ion transport, cellproliferation, Insulin Growth Factor (“IGF-II”) release, and IGF-IIreceptor expression in osteoblasts. Effects on Insulin Growth Factor-I(“IGF-I”) and IGF-II have also been demonstrated in rat fracture callus.Stimulation of transforming growth factor beta (“TGF-β”) messenger RNA(“mRNA”) with PEMF in a bone induction model in a rat has been shown.Studies have also demonstrated upregulation of TGF-β mRNA by PEMF inhuman osteoblast-like cell line designated MG-63, wherein there wereincreases in TGF-β1, collagen, and osteocalcin synthesis. PEMFstimulated an increase in TGF-β1 in both hypertrophic and atrophic cellsfrom human non-union tissue. Further studies demonstrated an increase inboth TGF-β1 mRNA and protein in osteoblast cultures resulting from adirect effect of EMF on a calcium/calmodulin-dependent pathway.Cartilage cell studies have shown similar increases in TGF-β1 mRNA andprotein synthesis from EMF, demonstrating a therapeutic application tojoint repair. Various studies conclude that upregulation of growthfactor production may be a common denominator in the tissue levelmechanisms underlying electromagnetic stimulation. When using specificinhibitors, EMF can act through a calmodulin-dependent pathway. It hasbeen previously reported that specific PEMF and PRF signals, as well asweak static magnetic fields, modulate Ca²⁺ binding to CaM in a cell-freeenzyme preparation. Additionally, upregulation of mRNA for BMP2 and BMP4with PEMF in osteoblast cultures and upregulation of TGF-β1 in bone andcartilage with PEMF have been demonstrated.

However, prior art in this field does not use an induction apparatusthat is lightweight, portable, disposable, implantable, and configuredwith, integrated into, or attached to at least one of garments, fashionaccessories, footware, bandages, anatomical supports, an anatomicalwraps, apparel, cushions, mattresses, pads, wheelchairs, therapeuticbeds, therapeutic chairs, therapeutic and health maintenance devicessuch as vacuum assisted wound closure devices, mechanical and functionalelectrical stimulation devices and exercise devices, ultrasound, heat,cold, massage, and exercise.

Therefore, a need exists for an electromagnetic treatment inductionapparatus and a method for using same that is lightweight, portable,implantable, and can be disposable. A further need exists for anelectromagnetic treatment induction apparatus and method that can beused more effectively with miniaturized circuitry that optimallyconfigures electromagnetic waveforms to be inductively coupled withplant, animal, and human tissue, organs, cells, and molecules fortherapeutic treatment.

As mentioned above, by use of a substantially single voltage amplitudeenvelope with each PRF burst, one was limiting frequency components thatcould couple to relevant dielectric pathways in cells and tissue.

However, prior art in this field does not configure waveforms based upona ion/ligand binding transduction pathway. Prior art waveforms areinefficient since prior art waveforms apply unnecessarily high amplitudeand power to living tissues and cells, require unnecessarily longtreatment time, and cannot be generated by a portable device.

Therefore, a need exists for an apparatus and a method that moreeffectively modulates angiogenesis and other biochemical processes thatregulate tissue growth and repair, shortens treatment times, andincorporates miniaturized circuitry and light weight applicators thusallowing the apparatus to be portable and if desired disposable. Afurther need exists for an apparatus and method that more effectivelymodulates angiogenesis and other biochemical processes that regulatetissue growth and repair, shortens treatment times, and incorporatesminiaturized circuitry and light weight applicators that can beconstructed to be implantable.

Time-varying electromagnetic fields, comprising either rectangular,pseudo-rectangular, or both rectangular and pseudo-rectangularwaveforms, such as pulse modulated electromagnetic fields, andsinusoidal waveforms such as pulsed radio frequency fields ranging fromseveral Hertz to an about 15 to an about 40 MHz range, are clinicallybeneficial when used as an adjunctive therapy for a variety ofmusculoskeletal injuries and conditions.

However, prior art in this field does not use an induction apparatusthat delivers a signal according to a mathematical model, isprogrammable, lightweight, portable, disposable, implantable, andconfigured with, integrated into, or attached to at least one ofgarments, fashion accessories, footware, bandages, anatomical supports,an anatomical wraps, apparel, cushions, mattresses, pads, wheelchairs,therapeutic beds, therapeutic chairs, therapeutic and health maintenancedevices such as vacuum assisted wound closure devices, mechanical andfunctional electrical stimulation devices and exercise devices,ultrasound, heat, cold, massage, and exercise. A further need exists foran electromagnetic treatment induction apparatus and a method for usingsame that is lightweight, portable, implantable, and can be disposable.A further need exists for an electromagnetic treatment inductionapparatus and method having decreased power requirements andnon-invasive characteristics that allow an enhanced signal to beintegrated into surgical dressings, wound dressings, pads, seatcushions, mattress pads, shoes, and any other garment and structurejuxtaposed to living tissue and cells, even to be integral to creationof a garment to deliver an enhanced EMF signal to any body parts andthat delivers a signal according to a mathematical model and isprogrammable.

Prior art equipment in this field is bulky, not designed for outdooruse, and not self-contained.

Therefore, a need exists for an apparatus and a method that moreeffectively modulates biochemical processes that regulate hair and othercerebrofacial tissue growth and repair, shortens treatment times, andincorporates miniaturized circuitry and light weight applicators thusallowing the apparatus to be portable and if desired disposable. Afurther need exists for an apparatus and method that more effectivelymodulates biochemical processes that regulate hair and othercerebrofacial tissue growth and repair, shortens treatment times, andincorporates miniaturized circuitry and light weight applicators thatcan be constructed to be implantable.

Traumatic brain injury (hereinafter known as TBI) remains as one of theleading causes of morbidity and mortality for civilians and for soldierson the battlefield and is a major health and socio-economic problemthroughout the world. In currently deployed war-fighters, head injuries,the majority of which include the brain, account for 22% of all injuriesand 56% of those are classified as moderate to severe. In January 2008,the Department of Defense reported that over 5,500 soldiers had sufferedtraumatic brain injury caused by explosive weaponry, including suicidebombings, mines that explode on impact, and missiles. In addition to theimmediate needs of the wounded, traumatic brain injury may createlong-term or even permanent cognitive, motor, and sensory disabilitiesthat require ongoing support, rehabilitation, and treatment.

Additionally, traumatic brain injury is also a significant cause ofdeath in civilians. Epidemiological data indicate that in the US, atleast 1.4 to 2 million people are treated for traumatic brain injuryevery year, resulting in 56,000 deaths and 18,000 survivors sufferingfrom neurological impairment. Annual costs in the US are estimated at$60 billion. The World Health Organization projected that by 2020, roadtraffic accidents, a major cause of traumatic brain injury, will rankthird as a cause of the global burden of disease and disablement, behindonly ischemic heart disease and unipolar depression. Recently, thedemographics of traumatic brain injury have shifted to include morecases due to falls in middle-aged and older subjects. It is predictedthat there will be 5 million head injuries over the next decade and 30million worldwide.

Tissue damage from head injuries such as traumatic brain injurygenerally arises from the mechanical damage of the trauma event andsubsequent secondary physiological responses to the trauma event. Forexample, moderate to severe traumatic brain injury can producemechanical damage by direct trauma to brain tissue that can cause thedisruption of cell membranes and blood vessels, resulting in direct andischemic neuronal death. Then, secondary physiological responses such asinflammation and swelling can result in further damage and even death ofhealthy brain tissue. Importantly, even in the absence of directmechanical injury (i.e. diffuse brain trauma), such secondaryphysiological responses can still occur and result in injury to healthybrain tissue. For example, astrocytes and microglia often react to headinjury conditions and by secreting destructive cytokines (e.g. IL-1β,TNF-α, IFN-γ, and IL-6) as well as other inflammatory molecules, such asglutamate, reactive oxygen and nitrogen species, which, alone, or incombination, can be neurotoxic.

While the primary and immediate consequences of mechanical trauma toneurons cannot be undone, secondary pathological sequelae, specificallybrain swelling and inflammation, are situational candidates forintervention. The toll of neurological deficits and mortality from TBIcontinue in the military and private sectors and, to date, there are nowidely successful medical or surgical interventions to prevent neuronaldeath.

Current medical practice has attempted to use pharmaceuticals tomitigate and prevent tissue damage and injury resulting from secondaryphysiological responses of traumatic brain injury with little success.For example, intravenous, high-dose corticosteroids have beenadministered to reduce cerebral inflammation after traumatic braininjury, but several studies have demonstrated that steroids can beneurotoxic. In fact, results from a clinical randomized trial in 2005tested whether a high dose regimen of the steroid methylprednisolonesodium succinate (MPSS), administered within 8 hours after injury, wouldimprove survival after head injury. This trial was planned to randomize20,000 patients and was powered to detect a drop in mortality from 15%to 13%, a small, but important improvement in outcome. However, the dataand safety monitoring board halted the trial after half of the patientswere enrolled as it became apparent that MPSS significantly increasedmortality of severe injuries from 17.9% to 21.1% (p=0.0001).

The search for alternatives to improve morbidity and mortality fromtraumatic brain injury has not been fruitful. At least 21 multi-centerclinical trials, aimed to determine the clinical value of a range ofapproaches, from steroids to calcium and glutamate antagonists toantioxidants and anti-fibrinolytic agents and hypothermia were conductedfrom 1985 to 2006, but unfortunately none have demonstrated a convincingbenefit in the overall traumatic brain injury population. In spite ofextremely promising pre-clinical data and early phase trials, no agenthas yet been shown convincingly in a phase III trial to have clearbenefit in terms of improving functional outcome after traumatic braininjury. Importantly, a common problem in these pharmacologicalapproaches is that all of the candidate drugs had potential deleteriousside effects on non-target tissue. In fact, the development ofpharmaceutical agents for traumatic brain injury has all but ceased withincreasing reluctance of the pharmaceutical industry to sponsor thetesting of new candidate therapies as uncertainty remains regardingbenefit.

Given the absence of treatment options for head trauma, there is a needfor a therapy that can target and reduce secondary physiologicalresponses such as inflammation, swelling, and intracranial pressurewhile also promoting repair and regrowth in and around the injured area.While EMF treatments have been explored for a variety of uses, thepossible benefits of PEMF in treating or preventing neurological injuryand degenerative conditions such as TBI, subarachnoid hemorrhage, brainischemia, stroke, and Alzheimer's or Parkinson's Disease are relativelyunknown. This is in part due to the fact that the secondaryphysiological responses (e.g. inflammatory) in the central nervoussystem (CNS) differ from that of the periphery systems for which PEMF iscurrently used. Moreover, attention has been focused on pharmaceuticaltreatments until recently. Accordingly, embodiments of the presentinvention address this need and provide methods and devices using PEMFto treat patients suffering from neurological injury (such as traumaticbrain injury) and secondary physiological responses arising from thatinjury.

Transient elevations in cytosolic Ca²⁺, from external stimuli as simpleas changes in temperature and receptor activation, or as complex asmechanical disruption of tissue, will activate CaM. Once Ca²⁺ ions arebound, a conformational change will allow CaM bind to and activate anumber of key enzymes involved in cell viability and function, such asthe endothelial and neuronal constitutive nitric oxide synthases (cNOS);eNOS and nNOS, respectively. As a consequence, NO is rapidly produced,albeit in lower concentrations than the explosive increases in NOproduced by inducible NOS (iNOS), during the inflammatory response. Incontrast, these smaller, transient increases in NO produced byCa/CaM-binding will activate soluble guanylyl cyclase (sGC), which willcatalyze the formation of cyclic guanosine monophosphate (cGMP). TheCaM/NO/cGMP signaling pathway can rapidly modulate blood flow inresponse to normal physiologic demands, as well as to inflammation.Importantly, this same pathway will also rapidly attenuate expression ofcytokines such as interleukin-1beta (IL-1β), and iNOS and stimulateanti-apoptotic pathways in neurons. All of these effects are mediated bycalcium and cyclic nucleotides, which in turn regulate growth factorssuch as basic fibroblast growth factor (FGF-2) and vascular endothelialgrowth factor (VEGF), resulting in pleiotrophic effects on cellsinvolved in tissue repair and maintenance.

In general, inflammatory response in the brain differs from that inother organs. It is exemplified by a more modest and delayed recruitmentof leukocytes into the brain than into peripheral organs. Brainmicroglia, in contrast, are activated and release inflammatory mediatorsbeginning within minutes to hours after TBI. The mediators often expressneurotoxic and neuroprotective properties. For example, cytokines mayeither promote damage or support recovery processes; in some cases,cytokines, such as interleukin-6, may perform both functions.

This invention teaches that rapid intervention after traumatic head,cerebral and neural injury with electromagnetic fields configured torapidly modulate the biochemical signaling cascades animals and humansemploy in response to physical and chemical perturbations willsignificantly reduce the pathological consequences of such injuries,thereby reducing morbidity and the cost of health care.

Bone growth stimulator (hereinafter known as BGS) electromagnetic fieldsare now part of the standard armamentarium of orthopedic practiceworldwide for the treatment of recalcitrant bone fractures. Radiofrequency signals, originally developed for deep tissue heating(diathermy), were shown to produce biological effects when applied atnon-thermal levels using pulse-modulation techniques to produce pulsedradio frequency (hereinafter known as PRF) signals, which is a subsetfrequency band within PEMF. At the cellular level, numerous studiesdemonstrate that BGS, PRF and other electromagnetic field (hereinafterknown as EMF) signals modulate the release of growth factors andcytokines.

Stimulation of transforming growth factor beta (“TGF-b”) messenger RNA(“mRNA”) with EMF in a bone induction model in a rat has been shown.Studies have also demonstrated upregulation of TGF-b mRNA by PEMF inhuman osteoblast-like cell line designated MG-63, wherein there wereincreases in TGF-b1, collagen, and osteocalcin synthesis. EMF stimulatedan increase in TGF-b1 in both hypertrophic and atrophic cells from humannon-union tissue. Further studies demonstrated an increase in bothTGF-b1 mRNA and protein in osteoblast cultures resulting from a directeffect of EMF on a calcium/calmodulin-dependent pathway. Cartilage cellstudies have shown similar increases in TGF-b1 mRNA and proteinsynthesis from EMF, demonstrating a therapeutic application to jointrepair.

However, prior art in this field has not produced electromagneticsignals configured specifically to instantaneously accelerate theasymmetrical kinetics of the binding of intracellular ions to theirassociated buffers which regulate the biochemical signaling pathwaysliving systems employ in response to brain tissue ischemia from stroke,traumatic brain injury, head injury, cerebral injury, neurologicalinjury and neurodegenerative diseases. The result is that there are nodevices currently in use for clinical applications of electromagneticfields for the treatment of brain tissue ischemia from stroke, traumaticbrain injury, head injury, cerebral injury, neurological injury andneurodegenerative diseases.

Therefore, a need exists for an apparatus and a method that modulatesthe biochemical pathways that regulate animal and human tissue responseto brain tissue ischemia from stroke, traumatic brain injury, headinjury, cerebral injury, neurological injury and neurodegenerativediseases by configuring EMF signals specifically to accelerate theasymmetrical kinetics of ion binding to intracellular buffers whichregulate the relevant biochemical signaling pathways. Some embodimentsprovide for a method that employs electromagnetic fields for rapidtreatment of brain tissue ischemia from stroke, traumatic brain injury,head injury, cerebral injury, neurological injury and neurodegenerativediseases. In another embodiment, an apparatus incorporates miniaturizedcircuitry and light weight coil applicators or electrodes thus allowingthe apparatus to be low cost, portable and, if desired, disposable. Afurther need exists for an apparatus and method that incorporates theasymmetrical kinetics of ion binding to intracellular buffers toconfigure electromagnetic waveforms to increase the rate of ion bindingand enhance the biochemical signaling pathways living systems employ inresponse to brain tissue ischemia from stroke, traumatic brain injury,head injury, cerebral injury, neurological injury and neurodegenerativediseases, and incorporates miniaturized circuitry and light weightapplicators that can be constructed to be implantable.

SUMMARY OF THE DISCLOSURE

Various apparatus, methods, devices, and systems are described herein.The summary, FIGS., and detailed descriptions are set forth in ten parts(parts 1-10). Each part may be considered internally consistent,however, embodiments, ranges, features, elements, and illustrations fromone part may be used in combination (in whole or in part) withembodiments, ranges, features, elements, and illustrations from anotherpart or parts. Although there is some repetition in the FIGS. andlanguages in each of these parts, this disclosure is intended toillustrate different variations and embodiments of the devices, systems,and methods for electrically stimulating tissue to treat variousdisorders, as described in greater detail herein.

Part 1

Described herein are apparatus and methods for deliveringelectromagnetic signals to human, animal and plant target pathwaystructures such as molecules, cells, tissue and organs for therapeuticand prophylactic purposes. A preferred embodiment according to thepresent invention utilizes a Power Signal to Noise Ratio (“Power SNR”)approach to configure bioeffective waveforms and incorporatesminiaturized circuitry and lightweight flexible coils. Thisadvantageously allows a device that utilizes a Power SNR approach,miniaturized circuitry, and lightweight flexible coils, to be completelyportable and if desired to be constructed as disposable and if furtherdesired to be constructed as implantable.

Specifically, broad spectral density bursts of electromagneticwaveforms, configured to achieve maximum signal power within a bandpassof a biological target, are selectively applied to target pathwaystructures such as living organs, tissues, cells and molecules.Waveforms are selected using a unique amplitude/power comparison withthat of thermal noise in a target pathway structure. Signals comprisebursts of at least one of sinusoidal, rectangular, chaotic and randomwave shapes, have frequency content in a range of about 0.01 Hz to about100 MHz at about 1 to about 100,000 bursts per second, and have a burstrepetition rate from about 0.01 to about 1000 bursts/second. Peak signalamplitude at a target pathway structure such as tissue, lies in a rangeof about 1 μV/cm to about 100 mV/cm. Each signal burst envelope may be arandom function providing a means to accommodate differentelectromagnetic characteristics of healing tissue. A preferredembodiment according to the present comprises a 20 millisecond pulseburst comprising about 5 to about 20 microsecond symmetrical orasymmetrical pulses repeating at about 1 to about 100 kilohertz withinthe burst. The burst envelope is a modified 1/f function and is appliedat random repetition rates. A resulting waveform can be delivered viainductive or capacitive coupling.

It is an object of the present invention to configure a power spectrumof a waveform by mathematical simulation by using signal to noise ratio(“SNR”) analysis to configure an optimized, bioeffective waveform thencoupling the configured waveform using a generating device such as ultralightweight wire coils that are powered by a waveform configurationdevice such as miniaturized electronic circuitry.

It is another object of the present invention to evaluate Power SNR forany target pathway structure such as molecules, cells, tissues andorgans of plants, animals and humans using any input waveform, even ifthe electrical equivalents are non-linear as in a Hodgkin-Huxleymembrane model.

It is another object of the present invention to provide a method andapparatus for treating plants, animals and humans using electromagneticfields selected by optimizing a power spectrum of a waveform to beapplied to a chosen biochemical target pathway structure such as amolecule, cell, tissue and organ of a plant, animal, and human.

It is another object of the present invention to employ significantlylower peak amplitudes and shorter pulse duration. This can beaccomplished by matching via Power SNR, a frequency range in a signal tofrequency response and sensitivity of a target pathway structure such asa molecule, cell, tissue, and organ, of plants, animals and humans.

Part 2

An electromagnetic treatment induction apparatus and a method for usingsame for therapeutic treatment of living tissues and cells byinductively coupling optimally configured waveforms to alter the livingtissues and cells' interaction with their electromagnetic environment.

According to an embodiment of the present invention, by treating aselectable body region with a flux path comprising a succession of EMFpulses having a minimum width characteristic of at least about 0.01microseconds in a pulse burst envelope having between about 1 and about100,000 pulses per burst, in which a voltage amplitude envelope of saidpulse burst is defined by a randomly varying parameter in whichinstantaneous minimum amplitude thereof is not smaller than the maximumamplitude thereof by a factor of one ten-thousandth. The pulse burstrepetition rate can vary from about 0.01 to about 10,000 Hz. Amathematically definable parameter can also be employed to define anamplitude envelope of said pulse bursts.

By increasing a range of frequency components transmitted to relevantcellular pathways, access to a large range of biophysical phenomenaapplicable to known healing mechanisms, including enhanced enzymeactivity and growth factor and cytokine release, is advantageouslyachieved.

According to an embodiment of the present invention, by applying arandom, or other high spectral density envelope, to a pulse burstenvelope of mono- or bi-polar rectangular or sinusoidal pulses whichinduce peak electric fields between 10⁻⁶ and 10 volts per centimeter(V/cm), a more efficient and greater effect can be achieved onbiological healing processes applicable to both soft and hard tissues inhumans, animals and plants. A pulse burst envelope of higher spectraldensity can advantageously and efficiently couple to physiologicallyrelevant dielectric pathways, such as, cellular membrane receptors, ionbinding to cellular enzymes, and general transmembrane potential changesthereby modulating angiogenesis and neovascularization.

By advantageously applying a high spectral density voltage envelope as amodulating or pulse-burst defining parameter, power requirements forsuch modulated pulse bursts can be significantly lower than that of anunmodulated pulse. This is due to more efficient matching of thefrequency components to the relevant cellular/molecular process.Accordingly, the dual advantages of enhanced transmitting dosimetry torelevant dielectric pathways and of decreasing power requirements areachieved.

A preferred embodiment according to the present invention comprisesabout 0.1 to about 100 millisecond pulse burst comprising about 1 toabout 200 microsecond symmetrical or asymmetrical pulses repeating atabout 0.1 to about 100 kilohertz within the burst. The burst envelope isa modified 1/f function and is applied at random repetition ratesbetween about 0.1 and about 1000 Hz. Fixed repetition rates can also beused between about 0.1 Hz and about 1000 Hz. An induced electric fieldfrom about 0.001 mV/cm to about 100 mV/cm is generated. Anotherembodiment according to the present invention comprises an about 0.01millisecond to an about 10 millisecond burst of high frequencysinusoidal waves, such as 27.12 MHz, repeating at about 1 to about 100bursts per second. An induced electric field from about 0.001 mV/cm toabout 100 mV/cm is generated. Resulting waveforms can be delivered viainductive or capacitive coupling.

It is another object of the present invention to provide anelectromagnetic method of treatment of living cells and tissuescomprising a broad-band, high spectral density electromagnetic field.

It is a further object of the present invention to provide anelectromagnetic method of treatment of living cells and tissuescomprising amplitude modulation of a pulse burst envelope of anelectromagnetic signal that will induce coupling with a maximum numberof relevant EMF-sensitive pathways in cells or tissues.

It is an object of the present invention to configure a power spectrumof a waveform by mathematical simulation by using signal to noise ratio(“SNR”) analysis to configure a waveform optimized to modulateangiogensis and neovascualarization then coupling the configuredwaveform using a generating device such as ultra lightweight wire coilsthat are powered by a waveform configuration device such as miniaturizedelectronic circuitry.

It is an object of the present invention to provide lightweight flexiblecoils, that can be placed in at least one of garments, fashionaccessories, footware, bandages, anatomical supports, an anatomicalwraps, apparel, cushions, mattresses, pads, wheelchairs, therapeuticbeds, therapeutic chairs, therapeutic and health maintenance devicessuch as vacuum assisted wound closure devices, mechanical and functionalelectrical stimulation devices and exercise devices and dressings todeliver the optimum dose of non-invasive pulsed electromagnetictreatment configured as shown above, for enhanced repair and growth ofliving tissue in animals, humans and plants.

It is another object of the present invention to provide multiple coils,delivering a waveform configured by SNR/Power analysis of a targetpathway, to increase area of treatment coverage.

It is another object of the present invention to provide multiple coilsthat are simultaneously driven or that are sequentially driven such asmultiplexed, with the same or different optimally configured waveformsas shown above.

It is a further object of the present invention to provide flexible,lightweight coils that focus the EMF signal to the affected tissue byincorporating the coils, delivering a waveform configured by SNR/Poweranalysis of a target pathway, into ergonomic support garments.

It is yet a further object of the present invention to utilizeconductive thread to create daily wear, and exercise and sports garmentshaving integrated coils, delivering a waveform configured by SNR/Poweranalysis of a target pathway, positioned in proximity to an anatomicaltarget.

It is yet a further object of the present invention to utilizelightweight flexible coils or conductive thread to deliver the EMFsignal to affected tissue by incorporating such coils or conductivethreads as an integral part of various types of bandages, such as,compression, elastic, cold compress and hot compress and delivering awaveform configured by SNR/Power analysis of a target pathway.

It is another object of the present invention to employ several coils,delivering a waveform configured by SNR/Power analysis of a targetpathway, to increase EMF coverage area.

It is another object of the present invention to construct a coil,delivering a waveform configured by SNR/Power analysis of a targetpathway, using conductive thread.

It is another object of the present invention to construct a coil,delivering a waveform configured by SNR/Power analysis of a targetpathway, using fine flexible conductive wire.

It is another object of the present invention to supply the same ordifferent waveforms configured by SNR/Power analysis of a targetpathway, simultaneously or sequentially to single or multiple coils.

It is yet a further object of the present invention to incorporate atleast one coil in a surgical wound dressing to apply an enhanced EMFsignal non-invasively and non-surgically, the surgical wound dressing tobe used in combination with standard wound treatment.

It is another object of the present invention to construct the coilsdelivering a waveform configured by SNR/Power analysis of a targetpathway, for easy attachment and detachment to dressings, garments andsupports by using an attachment means such as Velcro, an adhesive andany other such temporary attachment means.

It is another object of the present invention to provide coilsdelivering a waveform configured by SNR/Power analysis of a targetpathway, that are integrated with therapeutic beds, therapeutic chairs,and wheelchairs.

It is another object of the present invention to provide coilsdelivering a waveform configured by SNR/Power analysis of a targetpathway, that are integrated with various therapy surfaces, such aspressure relieving, inflatable, fluid, visco-elastic and air fluidizedbed and other support surfaces.

It is another object of the present invention to provide coilsdelivering a waveform configured by SNR/Power analysis of a targetpathway that are integrated with therapeutic seat cushions such asinflatable, fluidized, foam cushions.

It is another object of the present invention to provide coilsdelivering a waveform configured by SNR/Power analysis of a targetpathway, that are integrated with at least one of therapeutic mattressoverlays, sheets, blankets, pillows, pillow cases, and therapeuticdevices that can apply steady or intermittent pressure such as airclearance vests.

It is another object of the present invention to provide for theinclusion of a flux path to any therapeutic surface, structure, ordevice to enhance the effectiveness of such therapeutic surfaces,structures or devices by delivering a waveform configured by SNR/Poweranalysis of a target pathway.

It is another object of the present invention to incorporate coilsdelivering a waveform configured by SNR/Power analysis of a targetpathway, in footware such as shoes.

It is another object of the present invention to integrate at least onecoil delivering a waveform configured by SNR/Power analysis of a targetpathway, with a therapeutic surface, structure or device to enhance theeffectiveness of such therapeutic surface, structure or device.

Part 3

Also described herein are apparatus and methods for electromagnetictreatment of living tissues and cells by altering their interaction withtheir electromagnetic environment.

It is an object of the present invention to provide modulation ofelectromagnetically sensitive regulatory processes at the cell membraneand at junctional interfaces between cells.

It is another object of the present invention to provide increased bloodflow to affected tissues by modulating angiogenesis andneovascualarization.

It is another object of the present invention to provide increased bloodflow to enhance viability, growth, and differentiation of implantedcells, such as stem cells, tissues and organs.

It is another object of the present invention to provide increased bloodflow in cardiovascular diseases by modulating angiogenesis andneovascualarization.

It is another object of the present invention to improve micro-vascularblood perfusion and reduced transudation.

It is another object of the present invention to provide a treatment ofmaladies of the bone and other hard tissue by modulating angiogenesisand neovascularization.

It is a still further object of the present invention to provide atreatment of edema and swelling of soft tissue by increased blood flowthrough modulation of angiogenesis and neovascularization.

It is another object of the present invention to provide anelectromagnetic method of treatment of living cells and tissues that canbe used for repair of damaged soft tissue.

It is yet another object of the present invention to increase blood flowto damaged tissue by modulation of vasodilation and stimulatingneovascularization.

It is a yet further object of the present invention to provide anapparatus for modulation of angiogenesis and neovascularization that canbe operated at reduced power levels and still possess benefits ofsafety, economics, portability, and reduced electromagneticinterference.

It is another object of the present invention to modulate angiogenesisand neovascularization by evaluating Power SNR for any target pathwaystructure such as molecules, cells, tissues and organs of plants,animals and humans using any input waveform, even if electricalequivalents are non-linear as in a Hodgkin-Huxley membrane model.

It is another object of the present invention to provide a method andapparatus for treating plants, animals and humans using electromagneticfields selected by optimizing a power spectrum of a waveform to beapplied to a biochemical target pathway structure to enable modulationof angiogenesis and neovascularization within molecules, cells, tissuesand organs of a plant, animal, and human.

It is another object of the present invention to significantly lowerpeak amplitudes and shorter pulse duration. This can be accomplished bymatching via Power SNR, a frequency range in a signal to frequencyresponse and sensitivity of a target pathway structure such as amolecule, cell, tissue, and organ, of plants, animals and humans toenable modulation of angiogenesis and neovascularization.

Part 4

The present invention relates to accelerating wound repair of livingtissues, cells and molecules by providing a therapeutic, prophylacticand wellness apparatus and method for non-invasive pulsedelectromagnetic treatment to enhance condition, repair and growth ofliving tissue in animals, humans and plants. This beneficial methodoperates to selectively change a bio-electromagnetic environmentassociated with cellular and tissue environments by usingelectromagnetic means such as EMF generators and applicator heads. Anembodiment according to the present invention comprises introducing aflux path to a selectable body region, comprising a succession of EMFpulses having a minimum width characteristic of at least 0.01microseconds in a pulse burst envelope having between 1 and 100,000pulses per burst, in which a voltage amplitude envelope of said pulseburst is defined by a randomly varying parameter in which aninstantaneous minimum amplitude thereof is not smaller than a maximumamplitude thereof by a factor of one ten thousandth. Further, therepetition rate of such pulse bursts may vary from 0.01 to 10,000 Hertz.A mathematically definable parameter satisfying SNR and/or Power SNRdetectability requirements in a target structure is employed to definethe configuration of the pulse bursts.

It is another object of the present invention to provide a method oftreating living cells and tissue by electromagnetically modulatingsensitive regulatory processes at a cell membrane and at junctionalinterfaces between cells, using waveforms configured to satisfy SNR andPower SNR detectability requirements in a target pathway structure.

Specifically, broad spectral density bursts of electromagneticwaveforms, configured to achieve maximum signal power within a bandpassof a biological target, are selectively applied to target pathwaystructures such as tissues, to enhance effectiveness of pharmacological,chemical, cosmetic and topical agents. Waveforms are selected using aunique amplitude/power comparison with that of thermal noise in a targetpathway structure. Signals comprise bursts of at least one ofsinusoidal, rectangular, chaotic and random wave shapes, have frequencycontent in a range of about 0.01 Hz to about 100 MHz at about 1 to about100,000 bursts per second, and have a burst repetition rate from about0.01 to about 1000 bursts/second. Peak signal amplitude at a targetpathway structure such as organs, cells, tissues, and molecules, lies ina range of about 1 μV/cm to about 100 mV/cm. Each signal burst envelopemay be a random function providing a means to accommodate differentelectromagnetic characteristics of enhancing bioeffective processes. Apreferred embodiment according to the present invention comprises about0.1 to about 100 millisecond pulse burst comprising about 1 to about 200microsecond symmetrical or asymmetrical pulses repeating at about 0.1 toabout 100 kilohertz within the burst. The burst envelope is a modified1/f function and is applied at random repetition rates between about 0.1and about 1000 Hz. Fixed repetition rates can also be used between about0.1 Hz and about 1000 Hz. An induced electric field from about 0.001mV/cm to about 100 mV/cm is generated. Another embodiment according tothe present invention comprises an about 0.01 millisecond to an about 10millisecond burst of high frequency sinusoidal waves, such as 27.12 MHz,repeating at about 1 to about 100 bursts per second. An induced electricfield from about 0.001 mV/cm to about 100 mV/cm is generated. Resultingwaveforms can be delivered via inductive or capacitive coupling.

It is another object of the present invention to provide electromagnetictreatment for wound repair having a broad-band, high spectral densityelectromagnetic field configured according to at least one of SNR andPower SNR.

It is another object of the present invention to accelerate wound repairby configuring a power spectrum of a waveform by mathematical simulationby using signal to noise ratio (“SNR”) analysis to configure a waveformoptimized to modulate angiogensis and neovascualarization, then couplingthe configured waveform using a generating device such as ultralightweight wire coils that are powered by a waveform configurationdevice such as miniaturized electronic circuitry.

It is another object of the present invention to modulate angiogenesisand neovascularization by evaluating Power SNR at any target pathwaystructure such as molecules, cells, tissues and organs to acceleratewound repair by using any input waveform, even if electrical equivalentsare non-linear as in a Hodgkin-Huxley membrane model.

It is another object of the present invention to provide an apparatusthat incorporates use of Power SNR in which amplitude modulation of thepulse burst envelope of the electromagnetic signal will induce couplingwith a maximum number of relevant EMF-sensitive pathways in cells andtissues to enhance wound repair in humans, animals and plants.

It is another object of the present invention to provide a method andapparatus for enhancing wound repair using electromagnetic fieldsselected by optimizing a power spectrum of a waveform to be applied to abiochemical target pathway structure to enable modulation ofangiogenesis and neovascularization within molecules, cells, tissues andorgans.

It is another object of the present invention to significantly lowerpeak amplitudes and shorter pulse duration by matching via Power SNR, afrequency range in a signal to frequency response and sensitivity of atarget pathway structure such as a molecule, cell, tissue, and organthereby enabling modulation of angiogenesis and neovascularization foraccelerating wound repair.

It is another object of the invention to provide a method of enhancingsoft tissue and hard tissue repair.

It is another object of the invention to provide a method of increasingblood flow to affected tissues by modulating angiogenesis.

It is another object of the invention to provide an improved method ofincreasing blood flow to enhance the viability and growth ordifferentiation of implanted cells, tissues and organs.

It is another object of the invention to provide an improved method ofincreasing blood flow in cardiovascular diseases by modulatingangiogenesis.

It is another object of the invention to provide beneficialphysiological effects through improvement of micro-vascular bloodperfusion and reduced transudation.

It is another object of the invention to provide an improved method oftreatment of maladies of the bone and other hard tissue.

It is a still further object of the invention to provide an improvedmeans of the treatment of edema and swelling of soft tissue.

It is another object to provide a means of repair of damaged softtissue.

It is yet another object to provide a means of increasing blood flow todamaged tissue by modulation of vasodilation and stimulatingneovascularization.

It is yet another object to enhance healing of post-surgical wounds byreducing the inflammatory phase and modulating growth factor release.

It is yet another object of the instant invention to reduce theinflammatory phase post-cosmetic surgery.

It is yet another object of the instant invention to reduce or eliminatethe post-surgical complications of breast augmentation, such as capsularcontractions.

It is yet another object of the instant invention to reducepost-surgical pain, edema and discoloration.

It is yet a further object of the present invention to treat chronicwounds such as diabetic ulcers, venous stasis ulcers, pressure sores andany non-healing wound with EMF signals configured according to anembodiment of the present invention.

It is a yet a further object to provide apparatus for use of anelectromagnetic method of the character indicated, wherein operation ofthe apparatus can proceed at reduced power levels as compared to thoseof related methods known in electromedicine and respective biofieldtechnologies, with attendant benefits of safety, economics, portability,and reduced electromagnetic interference.

It is a further object of the present invention to provide a method fortreatment to enhance wellness.

It is a further object of the present invention to provide a method inwhich electromagnetic waveforms are configured according to SNR andPower SNR detectability requirements in a target pathway structure.

It is another object of the present invention to provide a method forelectromagnetic treatment comprising a broadband, high spectral densityelectromagnetic field.

It is another object of the present invention to provide a method ofenhancing soft tissue and hard tissue repair by using EMF.

It is another object of the present invention to provide a method toincrease blood flow to affected tissues by using electromagnetictreatment to modulate angiogenesis.

It is yet a further object of the present invention to provide a methodof treatment of chronic wounds such as diabetic ulcers, venous stasisulcers, pressure sores and any non-healing wound.

It is another object of the present invention to provide a method toincrease blood flow to regulate viability, growth, and differentiationof implanted cells, tissues and organs.

It is another object of the present invention to provide a method totreat cardiovascular diseases by modulating angiogensis and increasingblood flow.

It is another object of the present invention to provide a method toimprove micro-vascular blood perfusion and reduce transudation.

It is another object of the present invention to provide a method toincrease blood flow to treat maladies of bone and hard tissue.

It is another object of the present invention to provide a method toincrease blood flow to treat edema and swelling of soft tissue.

It is another object of the present invention to provide a method toincrease blood flow to repair damaged soft tissue.

It is another object of the present invention to provide a method toincrease blood flow to damaged tissue by modulation of vasodilation andstimulating neovascularization.

It is a further object of the present invention to provide anelectromagnetic treatment apparatus wherein the apparatus operates usingreduced power levels.

It is a yet further object of the present invention to provide anelectromagnetic treatment apparatus wherein the apparatus isinexpensive, portable, and produces reduced electromagneticinterference.

Part 5

The present invention relates to enhancing effectiveness ofpharmacological, chemical, cosmetic and topical agents used to treatliving tissues, cells and molecules by providing a therapeutic,prophylactic and wellness apparatus and method for non-invasive pulsedelectromagnetic treatment to enhance condition, repair and growth ofliving tissue in animals, humans and plants. This beneficial methodoperates to selectively change a bio-electromagnetic environmentassociated with cellular and tissue environments by usingelectromagnetic means such as EMF generators and applicator heads. Anembodiment according to the present invention comprises introducing aflux path to a selectable body region, comprising a succession of EMFpulses having a minimum width characteristic of at least 0.01microseconds in a pulse burst envelope having between 1 and 100,000pulses per burst, in which a voltage amplitude envelope of said pulseburst is defined by a randomly varying parameter in which aninstantaneous minimum amplitude thereof is not smaller than a maximumamplitude thereof by a factor of one ten thousandth. Further, therepetition rate of such pulse bursts may vary from 0.01 to 10,000 Hertz.A mathematically definable parameter satisfying SNR and/or Power SNRdetectability requirements in a target structure is employed to definethe configuration of the pulse bursts. Mathematically defined parametersare selected by considering the dielectric properties of the targetpathway structure, and the ratio of the induced electric field amplitudewith respect to voltage due to thermal noise or other baseline cellularactivity.

It is another object of the present invention to provide a method oftreating living cells and tissue by electromagnetically modulatingsensitive regulatory processes at a cell membrane and at junctionalinterfaces between cells, using waveforms configured to satisfy SNR andPower SNR detectability requirements in a target pathway structure.

It is another object of the present invention to enhance effectivenessof pharmacological, chemical, cosmetic and topical agents by configuringa power spectrum of a waveform by mathematical simulation by usingsignal to noise ratio (“SNR”) analysis to configure a waveform optimizedto modulate angiogensis and neovascualarization, then coupling theconfigured waveform using a generating device such as ultra lightweightwire coils that are powered by a waveform configuration device such asminiaturized electronic circuitry.

It is another object of the present invention to modulate angiogenesisand neovascularization by evaluating Power SNR at any target pathwaystructure such as molecules, cells, tissues and organs to enhanceeffectiveness of pharmacological, chemical, cosmetic and topical agents,by using any input waveform, even if electrical equivalents arenon-linear as in a Hodgkin-Huxley membrane model.

It is another object of the present invention to provide an apparatusthat incorporates use of Power SNR to regulate and adjustelectromagnetic therapy treatment to enhance effectiveness ofpharmacological, chemical, cosmetic and topical agents.

It is another object of the present invention to provide a method andapparatus for enhancing effectiveness of pharmacological, chemical,cosmetic and topical agents using electromagnetic fields selected byoptimizing a power spectrum of a waveform to be applied to a biochemicaltarget pathway structure to enable modulation of angiogenesis andneovascularization within molecules, cells, tissues and organs.

It is another object of the present invention to significantly lowerpeak amplitudes and shorter pulse duration by matching via Power SNR, afrequency range in a signal to frequency response and sensitivity of atarget pathway structure such as a molecule, cell, tissue, and organthereby enabling modulation of angiogenesis and neovascularization forenhancing effectiveness of pharmacological, chemical, cosmetic andtopical agents.

It is a further object of the present invention to provide an apparatusfor application of electromagnetic waveforms, to be used in conjunctionwith pharmacological, chemical, cosmetic and topical agents applied to,upon or in human, animal and plant cells, organs, tissues and moleculesso that bioeffective processes of such compounds can be enhanced.

It is a further object of the present invention to provide a method toenhance effectiveness of pharmacological, chemical, cosmetic and topicalagents for therapeutic, prophylactic and wellness ends.

It is a further object of the present invention to provide a method fortreatment of organs, muscles, joints, skin and hair using EMF inconjunction with pharmacological, chemical, cosmetic and topical agentsto improve the agents' effectiveness.

It is a further object of the present invention to provide a method fortreatment of organs, muscles, joints, skin and hair using EMF inconjunction with pharmacological, chemical, cosmetic and topical agentsto enhance wellness.

It is a further object of the present invention to provide a method inwhich electromagnetic waveforms are configured according to SNR andPower SNR detectability requirements in a target pathway structure.

It is another object of the present invention to provide a method forelectromagnetic treatment comprising a broadband, high spectral densityelectromagnetic field.

It is another object of the present invention to provide a method ofenhancing soft tissue and hard tissue repair by using EMF in conjunctionwith pharmacological, chemical, cosmetic and topical agents.

It is another object of the present invention to provide a method toenhance effectiveness of pharmacological, chemical, cosmetic and topicalagents by increasing blood flow to affected tissues by usingelectromagnetic treatment to modulate angiogenesis.

It is another object of the present invention to provide a method toincrease blood flow for enhancing effectiveness of pharmacological,chemical, cosmetic and topical agents that regulate viability, growth,and differentiation of implanted cells, tissues and organs.

It is another object of the present invention to provide a method totreat cardiovascular diseases by modulating angiogensis and increasingblood flow to enhance effectiveness of pharmacological, chemical,cosmetic and topical agents.

It is another object of the present invention to provide a method thatincreases physiological effectiveness of pharmacological, chemical,cosmetic and topical agents by improving micro-vascular blood perfusionand reduced transudation.

It is another object of the present invention to provide a method toincrease blood flow to enhance effectiveness of pharmacological,chemical, cosmetic and topical agents used for treating maladies of boneand hard tissue.

It is another object of the present invention to provide a method toincrease blood flow to enhance effectiveness of pharmacological,chemical, cosmetic and topical agents used for treating edema andswelling of soft tissue.

It is another object of the present invention to provide a method toincrease blood flow to enhance effectiveness of pharmacological,chemical, cosmetic and topical agents used for repairing damaged softtissue.

It is another object of the present invention to provide a method toincrease blood flow to damaged tissue by modulation of vasodilation andstimulating neovascularization whereby enhanced effectiveness ofpharmacological, chemical, cosmetic and topical agents is achieved.

It is a further object of the present invention to provide anelectromagnetic treatment apparatus wherein the apparatus operates usingreduced power levels.

It is a yet further object of the present invention to provide anelectromagnetic treatment apparatus wherein the apparatus isinexpensive, portable, and produces reduced electromagneticinterference.

Part 6

An electromagnetic treatment induction apparatus integrated intotherapeutic and non-therapeutic devices and a method for using same fortherapeutic treatment of living tissues and cells by inductivelycoupling optimally configured waveforms to alter the living tissues andcells' interaction with their electromagnetic environment.

The lightweight flexible coils can be an integral portion of apositioning device such as surgical dressings, wound dressings, pads,seat cushions, mattress pads, shoes, wheelchairs, chairs, and any othergarment and structure juxtaposed to living tissue and cells. Byadvantageously integrating a coil into a positioning device therapeutictreatment can be provided to living tissue and cells in an inconspicuousand convenient manner.

Specifically, broad spectral density bursts of electromagneticwaveforms, configured to achieve maximum signal power within a bandpassof a biological target, are selectively applied to target pathwaystructures such as living organs, tissues, cells and molecules.Waveforms are selected using a unique amplitude/power comparison withthat of thermal noise in a target pathway structure. Signals comprisebursts of at least one of sinusoidal, rectangular, chaotic and randomwave shapes, have frequency content in a range of about 0.01 Hz to about100 MHz at about 1 to about 100,000 bursts per second, and have a burstrepetition rate from about 0.01 to about 1000 bursts/second. Peak signalamplitude at a target pathway structure such as tissue, lies in a rangeof about 1 μV/cm to about 100 mV/cm. Each signal burst envelope may be arandom function providing a means to accommodate differentelectromagnetic characteristics of healing tissue. A preferredembodiment according to the present invention comprises about 0.1 toabout 100 millisecond pulse burst comprising about 1 to about 200microsecond symmetrical or asymmetrical pulses repeating at about 0.1 toabout 100 kilohertz within the burst. The burst envelope is a modified1/f function and is applied at random repetition rates between about 0.1and about 1000 Hz. Fixed repetition rates can also be used between about0.1 Hz and about 1000 Hz. An induced electric field from about 0.001mV/cm to about 100 mV/cm is generated. Another embodiment according tothe present invention comprises an about 0.01 millisecond to an about 10millisecond burst of high frequency sinusoidal waves, such as 27.12 MHz,repeating at about 1 to about 100 bursts per second. An induced electricfield from about 0.001 mV/cm to about 100 mV/cm is generated. Resultingwaveforms can be delivered via inductive or capacitive coupling.

It is another object of the present invention to deliver a waveformconfigured by SNR/Power analysis of a target pathway structure, in aprogrammable manner for example according to a time-dose program, aseries of pulses, or some other sequence random or patterned.

It is another object of the present invention to generate a signal froma waveform configured by SNR/Power analysis of a target pathwaystructure, in a programmable manner for example according to a time-doseprogram, a series of pulses, or some other sequence random or patterned.

It is yet another object of the present invention to integrate at leastone coil delivering a waveform configured by Power SNR analysis of atarget pathway structure, with at least one of a therapeutic surface, atherapeutic structure, and a therapeutic device, to enhance theeffectiveness of the at least one of the therapeutic surface, thetherapeutic structure, and the therapeutic device, to prevent the lossand deterioration of cells and tissues.

It is yet another object of the present invention to integrate at leastone coil delivering a waveform configured by Power SNR analysis of atarget pathway structure, with at least one of a therapeutic surface, atherapeutic structure, and a therapeutic device, to enhance theeffectiveness of the at least one of the therapeutic surface, thetherapeutic structure, and the therapeutic device, to augment cell andtissue activity.

It is yet another object of the present invention to integrate at leastone coil delivering a waveform configured by Power SNR analysis of atarget pathway structure, with at least one of a therapeutic surface, atherapeutic structure, and a therapeutic device, to enhance theeffectiveness of the at least one of the therapeutic surface, thetherapeutic structure, and the therapeutic device, to increase cellpopulation.

It is yet another object of the present invention to integrate at leastone coil delivering a waveform configured by Power SNR analysis of atarget pathway structure, with at least one of a therapeutic surface, atherapeutic structure, and a therapeutic device, to enhance theeffectiveness of the at least one of the therapeutic surface, thetherapeutic structure, and the therapeutic device, to prevent neurondeterioration.

It is yet another object of the present invention to integrate at leastone coil delivering a waveform configured by Power SNR analysis of atarget pathway structure, with at least one of a therapeutic surface, atherapeutic structure, and a therapeutic device, to enhance theeffectiveness of the at least one of the therapeutic surface, thetherapeutic structure, and the therapeutic device, to increase neuronpopulation.

It is yet another object of the present invention to integrate at leastone coil delivering a waveform configured by Power SNR analysis of atarget pathway structure, with at least one of a therapeutic surface, atherapeutic structure, and a therapeutic device, to enhance theeffectiveness of the at least one of the therapeutic surface, thetherapeutic structure, and the therapeutic device, to preventdeterioration of adrenergic neurons in a cerebrofacial area.

It is yet another object of the present invention to integrate at leastone coil delivering a waveform configured by Power SNR analysis of atarget pathway structure, with at least one of a therapeutic surface, atherapeutic structure, and a therapeutic device, to enhance theeffectiveness of the at least one of the therapeutic surface, thetherapeutic structure, and the therapeutic device, to increaseadrenergic neuron population in a cerebrofacial area.

Part 7

An apparatus and a method for electromagnetic treatment of hair andother cerebrofacial molecules, cells, organs, tissue, ions, and ligandsby altering their interaction with their electromagnetic environment.

By increasing a range of frequency components transmitted to relevantcellular pathways, hair and other cerebrofacial tissue restoration isadvantageously achieved.

According to an embodiment of the present invention, by applying arandom, or other high spectral density envelope, to a pulse burstenvelope of mono- or bi-polar rectangular or sinusoidal pulses whichinduce peak electric fields between 10⁻⁸ and 10 volts per centimeter(V/cm), a more efficient and greater effect can be achieved onbiological healing processes applicable to both soft and hard tissues inhumans, animals and plants. A pulse burst envelope of higher spectraldensity can advantageously and efficiently couple to physiologicallyrelevant dielectric pathways, such as, cellular membrane receptors, ionbinding to cellular enzymes, and general transmembrane potential changesthereby growing, restoring and maintaining hair and other cerebrofacialtissue.

Specifically, broad spectral density bursts of electromagneticwaveforms, configured to achieve maximum signal power within a bandpassof a biological target, are selectively applied to target pathwaystructures such as hair and other cerebrofacial tissues. Waveforms areselected using a unique amplitude/power comparison with that of thermalnoise in a target pathway structure. Signals comprise bursts of at leastone of sinusoidal, rectangular, chaotic and random wave shapes, havefrequency content in a range of about 0.01 Hz to about 100 MHz at about1 to about 100,000 bursts per second, and have a burst repetition ratefrom about 0.01 to about 1000 bursts/second. Peak signal amplitude at atarget pathway structure such as hair and or cerebrofacial tissue, liesin a range of about 1 μV/cm to about 100 mV/cm. Each signal burstenvelope may be a random function providing a means to accommodatedifferent electromagnetic characteristics of healing tissue. A preferredembodiment according to the present invention comprises about 0.1 toabout 100 millisecond pulse burst comprising about 1 to about 200microsecond symmetrical or asymmetrical pulses repeating at about 0.1 toabout 100 kilohertz within the burst. The burst envelope is a modified1/f function and is applied at random repetition rates between about 0.1and about 1000 Hz. Fixed repetition rates can also be used between about0.1 Hz and about 1000 Hz. An induced electric field from about 0.001mV/cm to about 100 mV/cm is generated. Another embodiment according tothe present invention comprises an about 0.01 millisecond to an about 10millisecond burst of high frequency sinusoidal waves, such as 27.12 MHz,repeating at about 1 to about 100 bursts per second. An induced electricfield from about 0.001 mV/cm to about 100 mV/cm is generated. Resultingwaveforms can be delivered via inductive or capacitive coupling.

It is another object of the present invention to provide anelectromagnetic method of treatment of hair and other cerebrofacialtissues comprising a broad-band, high spectral density electromagneticfield.

It is a further object of the present invention to provide anelectromagnetic method of treatment of hair and other cerebrofacialtissues comprising amplitude modulation of a pulse burst envelope of anelectromagnetic signal that will induce coupling with a maximum numberof relevant EMF-sensitive pathways in cells or tissues.

It is another object of the present invention to provide enhanced hairand other cerebrofacial tissue growth and repair in individuals thathave experienced hair loss due to medical conditions such as psoriasis,and hair loss as a result of medication shock and usage.

It is another object of the present invention to provide an apparatusand method that may be used in conjunction with pharmacological andherbal agents, and in conjunction with standard physical therapy andmedical treatments.

It is another object of the present invention to provide enhanced hairand other cerebrofacial tissue growth and repair in conjunction withtopical and medication treatments.

It is another object of the present invention to provide aself-contained hair restoration and cerebrofacial condition apparatusthat can be portable, fashionable, and worn whenever and wherever anindividual so desires.

It is another object of the present invention to provide aself-contained hair restoration and cerebrofacial condition apparatusthat can be programmed to release electromagnetic therapy treatment at,at least one of specific and random time intervals.

It is a still further object of the present invention to provide aself-contained hair restoration and cerebrofacial condition apparatusfor use in any type of headware, for example a hat, sweatband, andflexible knit cap.

It is yet another object of the present invention to increase blood flowto damaged cerebrofacial tissue by modulation of vasodilation andstimulating neovascularization.

It is yet another object of the present invention to prevent the lossand deterioration of cells and tissues of any type in the cerebrofacialarea.

A further object of the present invention is to augment the activity ofcells and tissues in the cerebrofacial area.

Yet a further object of the present invention is to increase cellpopulation in the cerebrofacial area.

It is yet a further object of the present invention to prevent thedeterioration of neurons in the cerebrofacial area.

It is yet another object of the present invention to increase neuronpopulation in the cerebrofacial area.

It is yet a further object of the present invention to prevent thedeterioration of adrenergic neurons in the cerebrofacial area.

It is yet another object of the present invention to increase adrenergicneuron population in the cerebrofacial area.

It is a yet another object of the present invention to provide anapparatus for cerebrofacial conditions that modulates angiogenesis andneovascularization that can be operated at reduced power levels andstill possess benefits of safety, economics, portability, and reducedelectromagnetic interference.

It is an object of the present invention to configure a power spectrumof a waveform by mathematical simulation by using signal to noise ratio(“SNR”) analysis to configure a waveform optimized to modulateangiogensis and neovascualarization in a cerebrofacial area thencoupling the configured waveform using a generating device such as ultralightweight wire coils that are powered by a waveform configurationdevice such as miniaturized electronic circuitry.

It is another object of the present invention to modulate angiogenesisand neovascularization by evaluating Power SNR for any target pathwaystructure such as molecules, cells, tissues and organs in acerebrofacial area using any input waveform, even if electricalequivalents are non-linear as in a Hodgkin-Huxley membrane model.

It is another object of the present invention to provide aself-contained hair restoration and cerebrofacial apparatus thatincorporates use of Power SNR to regulate and adjust electromagnetictherapy treatment.

It is another object of the present invention to provide a method andapparatus for treating hair loss and other cerebrofacial conditionsoccurring in animals and humans using electromagnetic fields selected byoptimizing a power spectrum of a waveform to be applied to a biochemicaltarget pathway structure to enable modulation of angiogenesis andneovascularization within molecules, cells, tissues and organs in acerebrofacial area.

It is another object of the present invention to significantly lowerpeak amplitudes and shorter pulse duration. This can be accomplished bymatching via Power SNR, a frequency range in a signal to frequencyresponse and sensitivity of a target pathway structure such as amolecule, cell, tissue, and organ, in a cerebrofacial area to enablemodulation of angiogenesis and neovascularization.

Part 8

An embodiment according to the present invention comprises anelectromagnetic signal having a pulse burst envelope of spectral densityto efficiently couple to physiologically relevant dielectric pathways,such as cellular membrane receptors, ion binding to cellular enzymes,and general transmembrane potential changes. The use of a burst durationwhich is generally below 100 microseconds for each PRF burst, limits thefrequency components that could couple to the relevant dielectricpathways in cells and tissue. An embodiment according to the presentinvention increases the number of frequency components transmitted torelevant cellular pathways whereby access to a larger range ofbiophysical phenomena applicable to known healing mechanisms, includingenhanced second messenger release, enzyme activity and growth factor andcytokine release can be achieved. By increasing burst duration andapplying a random, or other envelope, to the pulse burst envelope ofmono-polar or bi-polar rectangular or sinusoidal pulses which inducepeak electric fields between 10⁻⁶ and 10 V/cm, a more efficient andgreater effect can be achieved on biological healing processesapplicable to both soft and hard tissues in humans, animals and plants.

Another embodiment according to the present invention comprises knowncellular responses to weak external stimuli such as heat, light, sound,ultrasound and electromagnetic fields. Cellular responses to suchstimuli result in the production of protective proteins, for example,heat shock proteins, which enhance the ability of the cell, tissue,organ to withstand and respond to such external stimuli. Electromagneticfields configured according to an embodiment of the present inventionenhance the release of such compounds thus advantageously providing animproved means to enhance prophylactic protection and wellness of livingorganisms. In certain ophthalmic diseases there are physiologicaldeficiencies and disease states that can have a lasting and deleteriouseffect on the proper functioning of the ophthalmic system. Thosephysiological deficiencies and disease states can be positively affectedon a non-invasive basis by the therapeutic application of waveformsconfigured according to an embodiment of the present invention. Inaddition, electromagnetic waveforms configured according to anembodiment of the present invention can have a prophylactic effect onthe ophthalmic system whereby a disease condition can be prevented, andif a disease condition already exists in its earliest stages, thatcondition can be prevented from developing into a more advanced state.

An example of an ophthalmic disease that can be positively affected byan embodiment according to the present invention, both on a chronicdisease as well on a prophylactic basis, is macular degeneration.Age-related macular degeneration (“ARMD”) is the most common cause ofirreversible vision loss those over the age of 60. Macular degenerationis a disorder of the retina, the light-sensitive inner lining of theback of the eye. There are a number of abnormalities associated with theterm “age-related macular degeneration.” They range from mild changeswith no decrease in vision to abnormalities severe enough to result inthe loss of all “straight ahead” vision. Macular degeneration does notcause total blindness because the remaining and undamaged parts of theretina around the macula continue to provide “side” vision.

There are two main types of macular degeneration, “dry” and “wet.” Withrespect to dry macular degeneration, aging causes the cells in theretina to become less efficient. Deposits of tissue, called drusen,appear under the retina which can be identified through visualexamination. A few small drusen may cause no decrease in vision.However, if too many large drusen develop, vision will decrease. Theapplication of electromagnetic waveforms configured according to anembodiment of the present invention can positively effect tissue presentin the retina and modify the propensity to form drusen, thereby havingan effect on the progression of dry macular degeneration. Conversely,wet macular degeneration is a function of leaking of the capillaries inthe layer of cells below the retina called the retinal pigmentepithelium. Electromagnetic waveforms configured according to anembodiment of the present invention, have proven to have a positiveeffect on circulatory vessels and other tissues which can lead to animprovement in the disease state of wet macular degeneration.

Another advantage of electromagnetic waveforms configured according toan embodiment of the present invention is that by applying a highspectral density voltage envelope as the modulating or pulse-burstdefining parameter, the power requirement for such increased durationpulse bursts can be significantly lower than that of shorter pulsebursts containing pulses within the same frequency range; this is due tomore efficient matching of the frequency components to the relevantcellular/molecular process. Accordingly, the dual advantages, ofenhanced transmitted dosimetry to the relevant dielectric pathways andof decreased power requirement are achieved.

The present invention relates to a therapeutically beneficial method ofand apparatus for non-invasive pulsed electromagnetic treatment forenhanced condition, repair and growth of living tissue in animals,humans and plants. This beneficial method operates to selectively changethe bioelectromagnetic environment associated with the cellular andtissue environment through the use of electromagnetic means such as PRFgenerators and applicator heads. An embodiment of the present inventionmore particularly includes the provision of a flux path, to a selectablebody region, of a succession of EMF pulses having a minimum widthcharacteristic of at least 0.01 microseconds in a pulse burst envelopehaving between 1 and 100,000 pulses per burst, in which a voltageamplitude envelope of said pulse burst is defined by a randomly varyingparameter in which the instantaneous minimum amplitude thereof is notsmaller than the maximum amplitude thereof by a factor of oneten-thousandth. Further, the repetition rate of such pulse bursts mayvary from 0.01 to 10,000 Hz. Additionally a mathematically-definableparameter can be employed in lieu of said random amplitude envelope ofthe pulse bursts.

By increasing a range of frequency components transmitted to relevantcellular pathways, access to a large range of biophysical phenomenaapplicable to known healing mechanisms, including enhanced secondmessenger release, enzyme activity and growth factor and cytokinerelease, is advantageously achieved.

Another advantage of an embodiment according to the present invention isthat by applying a high spectral density voltage envelope as amodulating or pulse-burst defining parameter, power requirements forsuch modulated pulse bursts can be significantly lower than that of anunmodulated pulse. This is due to more efficient matching of thefrequency components to the relevant cellular/molecular process.Accordingly, the dual advantages of enhanced transmitting dosimetry torelevant dielectric pathways and of decreasing power requirements areachieved.

A further object of the present invention is to integrate at least onecoil delivering a waveform configured by SNR/Power analysis of a targetpathway structure, with a therapeutic surface, structure or device toenhance the effectiveness of such therapeutic surface, structure ordevice to augment the activity of cells and tissues of any type in anyliving target area.

It is yet a further object of the present invention to provide animproved electromagnetic method of the beneficial treatment of livingcells and tissue by the modulation of electromagnetically sensitiveregulatory processes at the cell membrane and at junctional interfacesbetween cells.

A further object of the present invention is to provide a means for theuse of electromagnetic waveforms to cause a beneficial effect in thetreatment of ophthalmic diseases.

It is a further object of the present invention to provide improvedmeans for the prophylactic treatment of the ophthalmic system to improvefunction and to prevent or arrest diseases of the ophthalmic system.

It is another object to provide an electromagnetic treatment method ofthe above type having a broad-band, high spectral densityelectromagnetic field.

It is a further object of the present invention to provide a method ofthe above type in which amplitude modulation of the pulse burst envelopeof the electromagnetic signal will induce coupling with a maximum numberof relevant EMF-sensitive pathways in cells or tissues.

It is another object of the present invention to provide an improvedmethod of enhancing soft tissue and hard tissue repair.

It is another object of the present invention to provide an improvedmethod of increasing blood flow to affected tissues by modulatingangiogenesis.

It is another object of the present invention to provide an improvedmethod of increasing blood flow to enhance the viability and growth ordifferentiation of implanted cells, tissues and organs.

It is another object of the present invention to provide an improvedmethod of increasing blood flow in cardiovascular diseases by modulatingangiogenesis.

It is another object of the present invention to provide beneficialphysiological effects through improvement of micro-vascular bloodperfusion and reduced transudation.

It is another object of the present invention to provide an improvedmethod of treatment of maladies of the bone and other hard tissue.

It is a still further object of the present invention to provide animproved means of the treatment of edema and swelling of soft tissue.

It is a still further object of the present invention to provide animproved means to enhance second messenger release.

It is another object of the present invention to provide a means ofrepair of damaged soft tissue.

It is yet another object of the present invention to provide a means ofincreasing blood flow to damaged tissue by modulation of vasodilationand stimulating neovascularization.

It is a yet further object of the present invention to provide anapparatus that can operate at reduced power levels as compared to thoseof related methods known in electromedicine and respective biofieldtechnologies, with attendant benefits of safety, economics, portability,and

Part 9

The methods and apparatus according to present invention, comprisesdelivering electromagnetic signals to respiratory target pathwaystructures, such as respiratory molecules, respiratory cells,respiratory tissues, and respiratory organs for treatment ofinflammatory processes leading to excessive fibrous tissue formationsuch as scar tissue, associated with the inhalation of foreign particlesinto lung tissue. An embodiment according to the present inventionutilizes SNR and Power SNR approaches to configure bioeffectivewaveforms and incorporates miniaturized circuitry and lightweightflexible coils. This advantageously allows a device that utilizes theSNR and Power SNR approaches, miniaturized circuitry, and lightweightflexible coils to be completely portable and if desired to beconstructed as disposable.

An embodiment according to the present invention comprises anelectromagnetic signal having a pulse burst envelope of spectral densityto efficiently couple to physiologically relevant dielectric pathways,such as cellular membrane receptors, ion binding to cellular enzymes,and general transmembrane potential changes. The use of a burst durationwhich is generally below 100 microseconds for each PRF burst, limits thefrequency components that could couple to the relevant dielectricpathways in cells and tissue. An embodiment according to the presentinvention increases the number of frequency components transmitted torelevant cellular pathways whereby access to a larger range ofbiophysical phenomena applicable to known healing mechanisms, includingenhanced second messenger release, enzyme activity and growth factor andcytokine release can be achieved. By increasing burst duration andapplying a random, or other envelope, to the pulse burst envelope ofmono-polar or bi-polar rectangular or sinusoidal pulses which inducepeak electric fields between 10⁻⁶ and 10 V/cm, a more efficient andgreater effect can be achieved on biological healing processesapplicable to both soft and hard tissues in humans, animals and plants.

Another embodiment according to the present invention comprises knowncellular responses to weak external stimuli such as heat, light, sound,ultrasound and electromagnetic fields. Cellular responses to suchstimuli result in the production of protective proteins, for example,heat shock proteins, which enhance the ability of the cell, tissue,organ to withstand and respond to such external stimuli. Electromagneticfields configured according to an embodiment of the present inventionenhance the release of such compounds thus advantageously providing animproved means to enhance prophylactic protection and wellness of livingorganisms. In certain respiratory diseases there are physiologicaldeficiencies and disease states that can have a lasting and deleteriouseffect on the proper functioning of the respiratory system. Thosephysiological deficiencies and disease states can be positively affectedon a non-invasive basis by the therapeutic application of waveformsconfigured according to an embodiment of the present invention. Inaddition, electromagnetic waveforms configured according to anembodiment of the present invention can have a prophylactic effect onthe respiratory system whereby a disease condition can be prevented, andif a disease condition already exists in its earliest stages, thatcondition can be prevented from developing into a more advanced state.

An example of a respiratory disease that can be positively affected byan embodiment according to the present invention, both on a chronicdisease as well on a prophylactic basis, is inflammation in lung tissueresulting from inhalation of foreign particles that remain in lungtissue. Electromagnetic waveforms configured according to an embodimentof the present invention, have proven to have a positive effect oncirculatory vessels and other tissues which can lead to reducinginflammation that can lead to lung disease.

Another advantage of electromagnetic waveforms configured according toan embodiment of the present invention is that by applying a highspectral density voltage envelope as the modulating or pulse-burstdefining parameter, the power requirement for such increased durationpulse bursts can be significantly lower than that of shorter pulsebursts containing pulses within the same frequency range; this is due tomore efficient matching of the frequency components to the relevantcellular/molecular process. Accordingly, the dual advantages, ofenhanced transmitted dosimetry to the relevant dielectric pathways andof decreased power requirement are achieved.

The present invention relates to a therapeutically beneficial method ofand apparatus for non-invasive pulsed electromagnetic treatment forenhanced condition, repair and growth of living tissue in animals,humans and plants. This beneficial method operates to selectively changethe bioelectromagnetic environment associated with the cellular andtissue environment through the use of electromagnetic means such as PRFgenerators and applicator heads. An embodiment of the present inventionmore particularly includes the provision of a flux path, to a selectablebody region, of a succession of EMF pulses having a minimum widthcharacteristic of at least 0.01 microseconds in a pulse burst envelopehaving between 1 and 100,000 pulses per burst, in which a voltageamplitude envelope of said pulse burst is defined by a randomly varyingparameter in which the instantaneous minimum amplitude thereof is notsmaller than the maximum amplitude thereof by a factor of oneten-thousandth. Further, the repetition rate of such pulse bursts mayvary from 0.01 to 10,000 Hz. Additionally a mathematically-definableparameter can be employed in lieu of said random amplitude envelope ofthe pulse bursts.

Another advantage of an embodiment according to the present invention isthat by applying a high spectral density voltage envelope as amodulating or pulse-burst defining parameter, power requirements forsuch modulated pulse bursts can be significantly lower than that of anunmodulated pulse. This is due to more efficient matching of thefrequency components to the relevant cellular/molecular process.Accordingly, the dual advantages of enhanced transmitting dosimetry torelevant dielectric pathways and of decreasing power requirements areachieved.

Specifically, broad spectral density bursts of electromagneticwaveforms, configured to achieve maximum signal power within a bandpassof a biological target, are selectively applied to target pathwaystructures such as living organs, tissues, cells and molecules.Waveforms are selected using a novel amplitude/power comparison withthat of thermal noise in a target pathway structure. Signals comprisebursts of at least one of sinusoidal, rectangular, chaotic and randomwave shapes have frequency content in a range of 0.01 Hz to 100 MHz at 1to 100,000 bursts per second, with a burst duration from 0.01 to 100milliseconds, and a burst repetition rate from 0.01 to 1000bursts/second. Peak signal amplitude at a target pathway structure suchas tissue, lies in a range of 1 μV/cm to 100 mV/cm. Each signal burstenvelope may be a random function providing a means to accommodatedifferent electromagnetic characteristics of healing tissue. Preferablythe present invention comprises a 20 millisecond pulse burst, repeatingat 1 to 10 burst/second and comprising 5 to 200 microsecond symmetricalor asymmetrical pulses repeating at 0.1 to 100 kilohertz within theburst. The burst envelope is a modified 1/f function and is applied atrandom repetition rates. Fixed repetition rates can also be used betweenabout 0.1 Hz and about 1000 Hz. An induced electric field from about0.001 mV/cm to about 100 mV/cm is generated. Another embodimentaccording to the present invention comprises a 4 millisecond of highfrequency sinusoidal waves, such as 27.12 MHz, repeating at 1 to 100bursts per second. An induced electric field from about 0.001 mV/cm toabout 100 mV/cm is generated. Resulting waveforms can be delivered viainductive or capacitive coupling for 1 to 30 minute treatment sessionsdelivered according to predefined regimes by which PEMF treatment may beapplied for 1 to 12 daily sessions, repeated daily. The treatmentregimens for any waveform configured according to the instant inventionmay be fully automated. The number of daily treatments may be programmedto vary on a daily basis according to any predefined protocol.

In another aspect of the present invention, an electromagnetic method oftreatment of living cells and tissues comprising modulation ofelectromagnetically sensitive regulatory processes at a cell membraneand at junctional interfaces between cells is provided.

In another aspect of the present invention, multiple coils deliver awaveform configured by SNR/Power analysis of a target pathway structure,to increase area of treatment coverage.

In another aspect of the present invention, multiple coils that aresimultaneously driven or that are sequentially driven such asmultiplexed, deliver the same or different optimally configuredwaveforms as illustrated above.

In still another aspect of the present invention, flexible, lightweightcoils that focus the EMF signal to the affected tissue delivering awaveform configured by SNR/Power analysis of a target pathway structure,are incorporated into dressings and ergonomic support garments.

In a further aspect of the present invention, at least one coildelivering a waveform configured by SNR/Power analysis of a targetpathway structure, is integrated with a therapeutic surface, structureor device to enhance the effectiveness of such therapeutic surface,structure or device to augment the activity of cells and tissues of anytype in any living target area.

In yet a further aspect of the present invention, an improvedelectromagnetic method of the beneficial treatment of living cells andtissue by the modulation of electromagnetically sensitive regulatoryprocesses at the cell membrane and at junctional interfaces betweencells is provided.

In a further aspect of the present invention, a means for the use ofelectromagnetic waveforms to cause a beneficial effect in the treatmentof respiratory diseases is provided.

In a further aspect of the present invention, improved means for theprophylactic treatment of the respiratory system to improve function andto prevent or arrest diseases of the respiratory system is provided.

In another aspect of the present invention, an electromagnetic treatmentmethod of the above type having a broad-band, high spectral densityelectromagnetic field is provided.

In a further aspect of the present invention, a method of the above typein which amplitude modulation of the pulse burst envelope of theelectromagnetic signal will induce coupling with a maximum number ofrelevant EMF-sensitive pathways in cells or tissues is provided.

In another aspect of the present invention, an improved method ofenhancing soft tissue and hard tissue repair is provided.

In another aspect of the present invention, an improved method ofincreasing blood flow to affected tissues by modulating angiogenesis isprovided.

In another aspect of the present invention, an improved method ofincreasing blood flow to enhance the viability and growth ordifferentiation of implanted cells, tissues and organs is provided.

In another aspect of the present invention, an improved method ofincreasing blood flow in cardiovascular diseases by modulatingangiogenesis is provided.

In another aspect of the present invention, beneficial physiologicaleffects through improvement of micro-vascular blood perfusion andreduced transudation are provided.

In another aspect of the present invention, an improved method oftreatment of maladies of the bone and other hard tissue is provided.

In still further aspect of the present invention, an improved means ofthe treatment of edema and swelling of soft tissue is provided.

In a further aspect of the present invention, an improved means toenhance second messenger release is provided.

In another aspect of the present invention, a means of repair of damagedsoft tissue is provided.

In yet another aspect of the present invention, a means of increasingblood flow to damaged tissue by modulation of vasodilation andstimulating neovascularization is provided.

In yet a further aspect of the present invention, an apparatus that canoperate at reduced power levels as compared to those of related methodsknown in electromedicine and respective biofield technologies, withattendant benefits of safety, economics, portability, and reducedelectromagnetic interference is provided.

“About” for purposes of the invention means a variation of plus or minus0.1%.

“Respiratory” for purposes of the invention means any organs andstructures such as nose, nasal passages, nasopharynx, larynx, trachea,bronchi, lungs and airways in which gas exchange takes.

Part 10

The apparatus and method according to present invention, comprisedelivering electromagnetic signals to fibrous capsule formation andcapsular contracture target pathway structures, such as capsularmolecules, capsular cells, capsular tissues, and capsular organs foralleviation of the propensity of a capsule to compress or harden, forreduction of excessive fibrous capsule formation, and for reduction inexisting capsule involvement with a physical area of a body. Anembodiment according to the present invention utilizes SNR and Power SNRapproaches to configure bioeffective waveforms and incorporatesminiaturized circuitry and lightweight flexible coils. Thisadvantageously allows a device that utilizes the SNR and Power SNRapproaches, miniaturized circuitry, and lightweight flexible coils to becompletely portable and if desired to be constructed as disposable.

An apparatus comprising an electromagnetic signal generating means foremitting signals comprising bursts of at least one of sinusoidal,rectangular, chaotic, and random waveforms, having a frequency contentin a range of about 0.01 Hz to about 100 MHz at about 1 to about 100,000waveforms per second, having a burst duration from about 1 usec to about100 msec, and having a burst repetition rate from about 0.01 to about1000 bursts/second, wherein the waveforms are adapted to have sufficientsignal to noise ratio of at least about 0.2 in respect of a givenfibrous capsule formation and capsular contracture target pathwaystructure to modulate at least one of ion and ligand interactions inthat fibrous capsule formation and capsular contracture target pathwaystructure, wherein the signal to noise ratio is evaluated by calculatinga frequency response of the impedance of the target path structuredivided by a calculated frequency response of baseline thermalfluctuations in voltage across the target path structure, anelectromagnetic signal coupling means wherein the coupling meanscomprises at least one of an inductive coupling means and a capacitivecoupling means, connected to the electromagnetic signal generating meansfor delivering the electromagnetic signal to the fibrous capsuleformation and capsular contracture target pathway structure, and agarment wherein the electromagnetic signal generating means andelectromagnetic signal coupling means are incorporated into the garment.

An apparatus comprising a waveform configuration means for configuringat least one waveform to have sufficient signal to noise ratio or powersignal to noise ratio of at least about 0.2, to modulate at least one ofion and ligand interactions whereby the at least one of ion and ligandinteractions are detectable in a fibrous capsule formation and capsularcontracture target pathway structure above baseline thermal fluctuationsin voltage and electrical impedance at the fibrous capsule formation andcapsular contracture target pathway structure, wherein the signal tonoise ratio is evaluated by calculating a frequency response of theimpedance of the target path structure divided by a calculated frequencyresponse of baseline thermal fluctuations in voltage across the targetpath structure, a coupling device connected to the waveformconfiguration means by at least one connecting means for generating anelectromagnetic signal from the configured at least one waveform and forcoupling the electromagnetic signal to the fibrous capsule formation andcapsular contracture target pathway structure whereby the at least oneof ion and ligand interactions are modulated, and a garmentincorporating the waveform configuration means, the at least oneconnecting means, and the coupling device.

A method comprising establishing baseline thermal fluctuations involtage and electrical impedance at a fibrous capsule formation andcapsular contracture target pathway structure depending on a state ofthe fibrous capsule tissue, evaluating a signal to noise ratio bycalculating a frequency response of the impedance of the target pathwaystructure divided by a calculated frequency response of baseline thermalfluctuations in voltage across the target pathway structure, configuringat least one waveform to have sufficient signal to noise ratio of atleast about 0.2 to modulate at least one of ion and ligand interactionswhereby the at least one of ion and ligand interactions are detectablein the fibrous capsule formation and capsular contracture target pathwaystructure above the evaluated baseline thermal fluctuations in voltage,generating an electromagnetic signal from the configured at least onewaveform; and coupling the electromagnetic signal to the fibrous capsuleformation and capsular contracture target pathway structure using acoupling device.

Part 11

Described herein are devices, systems and methods for deliveringelectromagnetic signals and fields configured specifically to acceleratethe asymmetrical kinetics of the binding of intracellular ions to theirrespective intracellular buffers, to enhance the biochemical signalingpathways animals and humans employ to respond to brain tissue ischemiafrom stroke, traumatic brain injury, head injury, cerebral injury,neurological injury and neurodegenerative diseases.

One variation according to the present invention utilizes a repetitiveburst of arbitrary non-thermal EMF waveforms configured to maximize thebound concentration of intracellular ions at their associated molecularbuffers to enhance the biochemical signaling pathways living systemsemploy in response to brain tissue ischemia from stroke, traumatic braininjury, head injury, cerebral injury, neurological injury andneurodegenerative diseases. Non-thermal electromagnetic waveforms areselected first by choosing the ion and the intracellular bindingprotein, for example Ca²⁺ and CaM, among the many ion-buffercombinations within the living cell, which determines the frequencyrange within which the signal must have non-thermal frequency componentsof sufficient, but non-destructive, amplitude to accelerate the kineticsof ion binding. Signals comprise a pulse duration, random signalduration or carrier period which is less than half of the ion bound timeto increase the voltage in the target pathway so as to maximallyaccelerate ion binding to maximally modulate biochemical signalingpathways to enhance specific cellular and tissue responses to braintissue ischemia from stroke, traumatic brain injury, head injury,cerebral injury, neurological injury and neurodegenerative diseases.

In some variations, signals comprise bursts of at least one ofsinusoidal, rectangular, chaotic or random EMF wave shapes; have burstduration less than about 100 msec, with frequency content less thanabout 100 MHz, repeating at less than about 1000 bursts per second. Peaksignal amplitude in the ion-buffer binding pathway is less than about1000 V/m. Another embodiment comprises about a 1 to about a 50millisecond burst of radio frequency sinusoidal waves in the range ofabout 1 to about 100 MHz, incorporating radio frequencies in theindustrial, scientific and medical (hereinafter known as ISM) band, forexample 27.12 MHz, but it may be 6.78 MHz, 13.56 MHz or 40.68 MHz in theshort wave frequency band, repeating between about 0.1 and about 10bursts/sec. Such waveforms can be delivered via inductive coupling witha coil applicator or via capacitive coupling with electrodes inelectrochemical contact with the conductive outer surface of the target.

Some embodiments described provide for a waveform configuration thataccelerates the kinetics of Ca²⁺ binding to CaM, consisting of about a 1to about a 10 msec burst of between about 5 MHz to about 50 MHz in theISM band, repeating between about 1 and about 5 bursts/sec and inducinga peak electric field between about 1 and about 100 V/m, then couplingthe configured waveform using a generating device such as ultralightweight wire or printed circuit coils that are powered by a waveformconfiguration device such as miniaturized electronic circuitry.

Other embodiments described provide for a waveform configuration thataccelerates the kinetics of Ca²⁺ binding to CaM, consisting of about a 1to about a 10 msec burst of 27.12 MHz radio frequency sinusoidal waves,repeating between about 1 and about 5 bursts/sec and inducing a peakelectric field between about 1 and about 100 V/m, then coupling theconfigured waveform using a generating device such as ultra lightweightwire, printed circuit coils or conductive garments that are powered by awaveform configuration device such as miniaturized electronic circuitrywhich is programmed to apply the aforementioned waveform at fixed orvariable intervals, for example for 1 minute every 10 minutes, or for 10minutes every hour, or for any other regimen found to be beneficial fora prescribed treatment. Further embodiments provide for methods anddevices for applying electromagnetic waveforms to animals and humansthat accelerate the asymmetrical kinetics of the binding ofintracellular ions to their associated intracellular buffers, byconfiguring the waveforms to contain repetitive frequency components ofsufficient amplitude to maximize the bound concentration of theintracellular ion to its associated intracellular buffer, thereby toenhance the biochemical signaling pathways living tissue employ inresponse to brain tissue ischemia from stroke, traumatic brain injury,head injury, cerebral injury, neurological injury and neurodegenerativediseases.

Additional embodiments provide for methods and devices for applyingelectromagnetic waveforms to animals and humans which match theasymmetrical kinetics of the binding of Ca²⁺ to CaM by configuring thewaveforms to contain repetitive frequency components of sufficientamplitude to accelerate and increase the binding of Ca²⁺ to CaM, therebyenhancing the CaM-dependent nitric oxide (NO)/cyclic guanosinemonophosphate (cGMP) signaling pathway.

Further embodiments provide for electromagnetic waveform configurationsto contain repetitive frequency components of sufficient amplitude toaccelerate and increase the binding of Ca²⁺ to CaM, thereby enhancingthe CaM-dependent NO/cGMP signaling pathway to accelerate blood andlymph vessel dilation for relief of post-operative and post traumaticpain and edema.

Another aspect of the present invention is to configure electromagneticwaveforms to contain repetitive frequency components of sufficientamplitude to accelerate and increase the binding of Ca²⁺ to CaM, therebyenhancing the CaM-dependent NO/cGMP signaling pathway, or any othersignaling pathway, to enhance angiogenesis and microvascularization forhard and soft tissue repair.

A further aspect of the present invention is to configureelectromagnetic waveforms to contain repetitive frequency components ofsufficient amplitude to accelerate and increase the binding of Ca²⁺ toCaM, thereby enhancing the CaM-dependent NO/cGMP signaling pathway, orany other signaling pathway, to accelerate deoxyribonucleic acid(hereinafter known as DNA) synthesis by living cells.

Another aspect of the present invention is to configure electromagneticwaveforms to contain repetitive frequency components of sufficientamplitude to accelerate and increase the binding of Ca²⁺ to CaM, therebyenhancing the CaM-dependent NO/cGMP signaling pathway to modulate growthfactor release, such as basic fibroblast growth factor (bFGF), vascularendothelial growth factor (VGEF), bone morphogenic protein (BMP), or anyother growth factor production by living cells.

It is yet another aspect of the present invention to configureelectromagnetic waveforms to contain repetitive frequency components ofsufficient amplitude to accelerate and increase the binding of Ca²⁺ toCaM, thereby enhancing the CaM-dependent NO/cGMP signaling pathway tomodulate growth factor release, such as basic fibroblast growth factor(bFGF), vascular endothelial growth factor (VGEF), bone morphogenicprotein (BMP), or any other growth factor production by living cellsemploy in response to brain tissue ischemia from stroke, traumatic braininjury, head injury, cerebral injury, neurological injury andneurodegenerative diseases.

Another aspect of the present invention is to configure electromagneticwaveforms to contain repetitive frequency components of sufficientamplitude to accelerate and increase the binding of Ca²⁺ to CaM, therebyenhancing the CaM-dependent NO/cGMP signaling pathway, or any othersignaling pathway, to modulate cytokine, such as interleukin 1-beta(IL-1β), interleukin-6 (IL-6), or any other cytokine production byliving cells.

Another aspect of the present invention is to configure electromagneticwaveforms to contain repetitive frequency components of sufficientamplitude to accelerate and increase the binding of Ca²⁺ to CaM, therebyenhancing the CaM-dependent NO/cGMP signaling pathway, or any othersignaling pathway, to modulate cytokine, such as interleukin 1-beta(IL-1β), interleukin-6 (IL-6), or any other cytokine production byliving cells in response to brain tissue ischemia from stroke, traumaticbrain injury, head injury, cerebral injury, neurological injury andneurodegenerative diseases.

Another aspect of the present invention is to configure electromagneticwaveforms to contain repetitive frequency components of sufficientamplitude to accelerate and increase the binding of Ca²⁺ to CaM, therebyenhancing the CaM-dependent NO/cGMP signaling pathway, or any othersignaling pathway, to accelerate the production of extracellularproteins for tissue repair and maintenance.

It is another aspect of the present invention to configureelectromagnetic waveforms to contain repetitive frequency components ofsufficient amplitude to accelerate and increase the binding of Ca²⁺ toCaM, thereby enhancing the CaM-dependent NO/cyclic adenosinemonophosphate (cAMP) signaling pathway, or any other signaling pathway,to modulate cell and tissue differentiation.

It is yet another aspect of the present invention to configureelectromagnetic waveforms to contain repetitive frequency components ofsufficient amplitude to accelerate and increase the binding of Ca²⁺ toCaM, thereby enhancing the CaM-dependent NO/cAMP signaling pathway, orany other signaling pathway, to prevent or reverse neurodegeneration.

Another aspect of the present invention is to configure electromagneticwaveforms to contain frequency components of sufficient amplitude toaccelerate the binding of Ca²⁺ to CaM, thereby enhancing theCaM-dependent NO/cGMP signaling pathway to modulate heat shock proteinrelease from living cells.

Yet another aspect of the invention provides for a method for treating aneurological injury or condition in a patient in need thereof includingthe steps of generating a pulsed electromagnetic field from a pulsedelectromagnetic field source and applying the pulsed electromagneticfield in proximity to a target region affected by the neurologicalinjury or condition to reduce a physiological response to theneurological injury or condition. Optionally, in any of the precedingembodiments, the physiological response can be inflammation and/orincreased intracranial pressure.

Optionally, in any of the preceding embodiments, the method may alsoinclude monitoring the physiological response and continuing to applythe pulsed electromagnetic field until an acceptable level of thephysiological response is reached. Optionally, in any of the precedingembodiments, the physiological response can be increased intracranialpressure and the acceptable level is below about 20 mmHg.

In further variations, the method may include a pulsed electromagneticfield comprising a 2 msec burst of 27.12 MHz sinusoidal waves repeatingat 2 Hz. In other variations, the method may include a pulsedelectromagnetic field comprising a 3 msec burst of 27.12 MHz sinusoidalwaves repeating at 2 Hz. In further embodiments, the pulsedelectromagnetic field may comprise a 4 msec burst of 27.12 MHzsinusoidal waves repeating at 2 Hz.

A further aspect of the invention provides for a method for treating aneurological injury or condition in a patient in need thereof where themethod includes generating a first pulsed electromagnetic field from apulsed electromagnetic field source; applying the first pulsedelectromagnetic field in proximity to a target region affected by theneurological injury or condition to reduce a physiological response tothe neurological injury or condition for a first treatment interval;discontinuing the application of the first pulsed electromagnetic fieldfor an inter-treatment period greater than zero; and applying a secondpulsed electromagnetic field in proximity to the target region.Optionally, in any of the preceding embodiments, the first and secondpulsed electromagnetic fields are substantially the same.

Optionally, in any of the preceding embodiments, the method may includemonitoring the physiological response; and modifying the first pulsedelectromagnetic field to the second pulsed electromagnetic field inresponse to the monitoring step.

Moreover, optionally, in any of the preceding embodiments, the methodmay also include monitoring the physiological response; anddiscontinuing treatment once an acceptable level of the physiologicalresponse is reached.

Optionally, in any of the preceding embodiments, the method may alsoinclude attenuating inflammatory cytokines and growth factors at thetarget region by applying the first pulsed electromagnetic field or thesecond pulsed electromagnetic field to the target region.

Optionally, in any of the preceding embodiments, the method may alsoinclude accelerating the healing of the target region by applying thefirst pulsed electromagnetic field or the second pulsed electromagneticfield to the target region.

Furthermore, in other embodiments, applying the first pulsedelectromagnetic field in proximity to a target region affected by theneurological injury or condition to reduce a physiological response maycomprise reducing a concentration of IL-1β. In further embodiments, theneurological injury or condition may be a neurodegenerative disease.

In further embodiments, the neurological injury or condition is TBI.

Another aspect of the invention provides for a method for treating aneurological injury or condition in a patient in need thereof, themethod including generating a pulsed electromagnetic field from a pulsedelectromagnetic field source; and applying the pulsed electromagneticfield in proximity to a target brain region affected by the neurologicalinjury or condition to reduce a physiological response to theneurological injury or condition by modulating microglia activation inthe target brain region. In some embodiments, modulating microgliaactivation includes reducing microglia activation in the target brainregion.

Another aspect of the invention provides for a method of promotingneurological repair or growth following a neurological injury orcondition including placing a treatment coil of a self-contained,lightweight, and portable treatment apparatus externally to a targettreatment site in need of repair or development, wherein the treatmentapparatus comprises a conformable coil having one or more turns of wireand a control circuit; generating an electromagnetic field using thetreatment coil; delivering the electromagnetic field to the targettreatment site using the treatment coil; and reducing a physiologicalresponse to the neurological injury or condition.

Optionally, in any of the preceding embodiments, generating anelectromagnetic field comprises generating at least one burst ofsinusoidal, rectangular, chaotic, or random waveforms, having afrequency content in a range of about 0.01 Hz to about 10,000 MHz atabout 1 to about 100,000 bursts per second, having a burst duration fromabout 0.01 to about 1000 bursts per second, and having a burstrepetition rate from about 0.01 to about 1000 bursts/second.

Generating an electromagnetic field may comprise generating at least oneburst of sinusoidal, rectangular, chaotic, or random waveforms, having afrequency content in a range of about 0.01 Hz to about 10,000 MHz,having a burst duration from about 0.1 to about 100 msec, at a peakamplitude of 0.001 G to about 0.1 G, and having a burst repetition ratefrom about 0.01 to about 100 bursts/second.

Optionally, in any of the preceding embodiments, the method may alsoinclude delivering the electromagnetic field for a period of about 1minute to about 240 minutes.

Optionally, in any of the preceding embodiments, the physiologicalresponse can be a cognitive deficiency.

Optionally, in any of the preceding embodiments, the pulsedelectromagnetic field comprises about a 1 msec to about a 10 msec burstof 27.12 MHz sinusoidal waves repeating at about 1 Hz to about 10 Hz.

Optionally, in any of the preceding embodiments, the pulsedelectromagnetic field comprises an ISM carrier frequency modulated atabout a 1 msec to about a 10 msec burst repeating at about 1 Hz to about10 Hz.

Optionally, in any of the preceding embodiments, the pulsedelectromagnetic field comprises an ISM carrier or any other radiofrequency up to 10,000 GHz, configured to modulate a rhythm of aphysiological system.

Optionally, in any of the preceding embodiments, the physiologicalsystem is the central nervous system. Moreover, optionally, in any ofthe preceding embodiments, the physiological system is the peripheralnervous system. Additionally, optionally, in any of the precedingembodiments, the physiological system is the cardiac system.

Optionally, in any of the preceding embodiments, the physiologicalsystem is the pulmonary system.

Optionally, in any of the preceding embodiments, the pulsedelectromagnetic field comprises an ISM carrier or any other radiofrequency up to 10,000 GHz, configured to modulate a rhythm of aphysiological process

Optionally, in any of the preceding embodiments, the pulsedelectromagnetic field comprises an ISM carrier or any other radiofrequency up to 10,000 GHz, configured to modulate a rhythm of a brain.

Optionally, in any of the preceding embodiments, the pulsedelectromagnetic field comprises an ISM carrier or any other radiofrequency up to 10,000 GHz, configured to modulate a circadian rhythm.

Optionally, in any of the preceding embodiments, the pulsedelectromagnetic field comprises an ISM carrier frequency configured tomodulate quality of sleep.

Optionally, in any of the preceding embodiments, the pulsedelectromagnetic field is configured to modulate calmodulin-dependentsignaling in a biological system.

Optionally, in any of the preceding embodiments, the electromagneticfield comprises a waveform that produces an effect uponcalmodulin-dependent signaling in a biological system.

Optionally, in any of the preceding embodiments, the electromagneticfield comprises a waveform that modulates at least one biologicalsignaling pathway.

Optionally, in any of the preceding embodiments, the method may alsoinclude increasing a growth factor in the target region.

Optionally, in any of the preceding embodiments, increasing a growthfactor in the target region enhances angiogenesis.

Optionally, in any of the preceding embodiments, increasing a growthfactor in the target region enhances nervous tissue regeneration.

Optionally, in any of the preceding embodiments, the growth factor isselected from the group consisting of FGF-2, VEGF, and BMP.

Optionally, in any of the preceding embodiments, the pulsedelectromagnetic field comprises an ISM carrier or any other radiofrequency up to 10,000 GHz, configured to modulate a sleep pattern.

Optionally, in any of the preceding embodiments, the pulsedelectromagnetic field comprises an ISM carrier or any other radiofrequency up to 10,000 GHz, configured to modulate slow-wave sleep in asleep cycle to effect the production of human growth hormone. The aboveand yet other embodiments and advantages of the present invention willbecome apparent from the hereinafter set forth Brief Description of theDrawings and Detailed Description of the Invention.

“About” for purposes of the invention means a variation of plus or minus50%.

The above and yet other aspects and advantages of the present inventionwill become apparent from the hereinafter set forth Brief Description ofthe Drawings and Detailed Description of the Invention.

BRIEF DESCRIPTION OF THE DRAWINGS

Preferred embodiments of the present invention will be described belowin more detail, with reference to the accompanying drawings:

Part 1

FIG. 1 is a flow diagram of a method for electromagnetic treatment ofplant, animal, and human target pathway structures such as tissue,organs, cells, and molecules according to an embodiment of the presentinvention;

FIG. 2 is a view of control circuitry and electrical coils applied to aknee joint according to a preferred embodiment of the present invention;

FIG. 3 is a block diagram of miniaturized circuitry according to apreferred embodiment of the present invention;

FIG. 4A is a line drawing of a wire coil such as an inductor accordingto a preferred embodiment of the present invention;

FIG. 4B is a line drawing of a flexible magnetic wire according to apreferred embodiment of the present invention;

FIG. 5 depicts a waveform delivered to a target pathway structure suchas a molecule, cell, tissue or organ according to a preferred embodimentof the present invention;

FIG. 6 is a view of a positioning device such as a wrist supportaccording to a preferred embodiment of the present invention;

FIG. 7 is a graph illustrating maximally increased myosinphosphorylation for a PMRF signal configured according to an embodimentof the present invention; and

FIG. 8 is a graph illustrating a power consumption comparison between a60 Hz signal and a PEMF signal configured according to an embodiment ofthe present invention.

Part 2

FIG. 9 is a flow diagram of a method for using an electromagnetictreatment inductive apparatus according to an embodiment of the presentinvention;

FIG. 10 is a view of control circuitry according to a preferredembodiment of the present invention;

FIG. 11 is a block diagram of miniaturized circuitry according to apreferred embodiment of the present invention;

FIG. 12 depicts an electromagnetic treatment inductive apparatusintegrated into a hip, thigh, and lower back support garment accordingto a preferred embodiment of the present invention;

FIG. 13 depicts an electromagnetic treatment inductive apparatusintegrated into a head and face support garment according to a preferredembodiment of the present invention;

FIG. 14 depicts an electromagnetic treatment inductive apparatusintegrated into a surgical dressing on a human forearm according to apreferred embodiment of the present invention;

FIG. 15 depicts an electromagnetic treatment inductive apparatusintegrated into a mattress pad according to a preferred embodiment ofthe present invention;

FIG. 16A depicts an electromagnetic treatment inductive apparatusintegrated into a sock according to a preferred embodiment of thepresent invention;

FIG. 16B depicts an electromagnetic treatment inductive apparatusintegrated into a shoe according to a preferred embodiment of thepresent invention;

FIG. 17 depicts an electromagnetic treatment inductive apparatusintegrated into a therapeutic bed according to a preferred embodiment ofthe present invention; and

FIG. 18 depicts an electromagnetic treatment inductive apparatusintegrated into a chest garment according to a preferred embodiment ofthe present invention.

Part 3

FIG. 19 is a flow diagram of a electromagnetic treatment method forangiogenesis modulation of living tissues and cells according to anembodiment of the present invention;

FIG. 20 is a view of control circuitry according to a preferredembodiment of the present invention;

FIG. 21 is a block diagram of miniaturized circuitry according to apreferred embodiment of the present invention;

FIG. 22 depicts a waveform delivered to a angiogenesis andneovascularization target pathway structure according to a preferredembodiment of the present invention.

Part 4

FIG. 23 is a flow diagram of a method for accelerating wound repair inliving tissues, cells and molecules according to an embodiment of thepresent invention;

FIG. 24 is a view of control circuitry and electrical coils applied to aknee joint according to a preferred embodiment of the present invention;

FIG. 25 is a block diagram of miniaturized circuitry according to apreferred embodiment of the present invention;

FIG. 26A is a line drawing of a wire coil such as an inductor accordingto a preferred embodiment of the present invention;

FIG. 26B is a line drawing of a flexible magnetic wire according to apreferred embodiment of the present invention;

FIG. 27 depicts a waveform delivered to a target pathway structure suchas a molecule, cell, tissue or organ according to a preferred embodimentof the present invention;

FIG. 28 is a view of a positioning device such as a wrist supportaccording to a preferred embodiment of the present invention;

FIG. 29 is a view of a positioning device such as a mattress padaccording to a preferred embodiment of the present invention;

FIG. 30 is a view of a positioning device such as a chest garmentaccording to an embodiment of the present invention;

FIG. 31 is a graph illustrating maximally increased myosinphosphorylation for a PMRF signal configured according to an embodimentof the present invention.

Part 5

FIG. 32 is a flow diagram of a method for enhancing effectiveness ofpharmacological, chemical, cosmetic and topical agents used to treatliving tissues, cells and molecules according to an embodiment of thepresent invention;

FIG. 33 is a view of control circuitry and electrical coils applied to aknee joint according to a preferred embodiment of the present invention;

FIG. 34 is a block diagram of miniaturized circuitry according to apreferred embodiment of the present invention;

FIG. 35A is a line drawing of a wire coil such as an inductor accordingto a preferred embodiment of the present invention;

FIG. 35B is a line drawing of a flexible magnetic wire according to apreferred embodiment of the present invention;

FIG. 36 depicts a waveform delivered to a target pathway structure suchas a molecule, cell, tissue or organ according to a preferred embodimentof the present invention;

FIG. 37 is a view of a positioning device such as a wrist supportaccording to a preferred embodiment of the present invention;

FIG. 38 is a view of a positioning device such as a mattress padaccording to a preferred embodiment of the present invention;

FIG. 39 is a graph illustrating effects of increased burst durationaccording to an embodiment of the present invention; and

FIG. 40 is a graph illustrating an increase in skin blood perfusionachieved according to an embodiment of the present invention.

Part 6

FIG. 41 is a flow diagram of a electromagnetic therapeutic treatmentmethod for using coils integrated into a positioning device according toan embodiment of the present invention;

FIG. 42 is a view of an electromagnetic treatment apparatus according toa preferred embodiment of the present invention;

FIG. 43 is a block diagram of miniaturized circuitry according to apreferred embodiment of the present invention;

FIG. 44 depicts a waveform delivered to a target pathway structureaccording to a preferred embodiment of the present invention.

FIG. 45 is a bar graph illustrating PMF pre-treatment results;

FIG. 46 is a bar graph illustrating specific PMF signal results; and

FIG. 47 is a bar graph illustrating chronic PMF results.

Part 7

FIG. 48 is a flow diagram of a electromagnetic treatment method for hairrestoration and cerebrofacial conditions according to an embodiment ofthe present invention;

FIG. 49 is a view of an electromagnetic treatment apparatus for hairrestoration and cerebrofacial conditions according to a preferredembodiment of the present invention;

FIG. 50 is a block diagram of miniaturized circuitry according to apreferred embodiment of the present invention; and

FIG. 51 depicts a waveform delivered to a hair and cerebrofacial targetpathway structure according to a preferred embodiment of the presentinvention;

FIG. 52 is a bar graph illustrating various burst width results;

FIG. 53 is a bar graph illustrating specific PMF signal results; and

FIG. 54 is a bar graph illustrating chronic PMF results.

Part 8

FIG. 55 is a flow diagram of a electromagnetic treatment method fortreatment of the ophthalmic tissue area according to an embodiment ofthe present invention;

FIG. 56 is a view of an electromagnetic treatment apparatus forophthalmic tissue treatment according to a preferred embodiment of thepresent invention;

FIG. 57 is a block diagram of miniaturized circuitry according to apreferred embodiment of the present invention;

FIG. 58 is a block diagram of miniaturized circuitry according toanother embodiment of the present invention;

FIG. 59 depicts a waveform delivered to eye target pathway structureaccording to a preferred embodiment of the present invention;

FIG. 60 is a bar graph illustrating PMF pre-treatment results;

FIG. 61 is a bar graph illustrating specific PMF signal results; and

FIG. 62 is a bar graph illustrating chronic PMF results.

Part 9

FIG. 63 is a flow diagram of a method for altering an electromagneticenvironment of respiratory tissue according to an embodiment of thepresent invention;

FIG. 64 is a view of an electromagnetic apparatus for respiratory tissuetreatment according to an embodiment of the present invention;

FIG. 65 is a block diagram of miniaturized circuitry according to anembodiment of the present invention;

FIG. 66 depicts a waveform delivered to a respiratory target pathwaystructure according to an embodiment of the present invention;

FIG. 67 is a view of inductors placed in a vest according to anembodiment of the present invention;

FIG. 68 is a bar graph illustrating myosin phosphorylation for a PMFsignal configured according to an embodiment of the present invention;and

FIG. 69 is a bar graph illustrating SNR signal effectiveness in a cellmodel of inflammation.

Part 10

FIG. 70 is a flow diagram of a method for altering fibrous capsuleformation and capsular contracture according to an embodiment of thepresent invention;

FIG. 71 is a view of an apparatus for application of electromagneticsignals according to an embodiment of the present invention;

FIG. 72 is a block diagram of miniaturized circuitry according to anembodiment of the present invention;

FIG. 73 depicts a waveform delivered to a capsule formation and capsulecontracture target pathway structure according to an embodiment of thepresent invention;

FIG. 74 is a view of inductors placed in a vest according to anembodiment of the present invention;

FIG. 75 is a bar graph illustrating myosin phosphorylation for a PMFsignal configured according to an embodiment of the present invention;and

FIG. 76 is a bar graph illustrating SNR signal effectiveness in a cellmodel of inflammation.

Part 11

FIG. 77A is a schematic representation of the biological EMFtransduction pathway which is a representative target pathway of EMFsignals configured according to embodiments described.

FIG. 77B is a flow diagram of a method for treating a neurologicalcondition/injury according to an embodiment of the devices and methodsdescribed herein.

FIG. 78A is a block diagram of miniaturized circuitry for use with acoil applicator according to some embodiments described.

FIG. 78B illustrates a device for application of electromagnetic signalsaccording to an embodiment of the devices and methods described herein.

FIG. 78C illustrates a waveform delivered to a target pathway structureof a plant, animal or human, such as a molecule cell, tissue, organ, orpartial or entire organism, according to some embodiments described.

FIGS. 79A and 79B illustrates the effect of a PEMF treatment accordingto embodiments described on nitric oxide (NO) release from MN9D neuronalcell cultures.

FIG. 80 illustrates the effect of a PEMF treatment according toembodiments described on angiogenesis in thermal myocardial necrosis ina rat model.

FIG. 81 illustrates the effect of a PEMF treatment according toembodiments described on edema formation in a carrageenan-induced pawedema model of inflammation in the rat.

FIGS. 82A-82C illustrate the effect of a PEMF treatment according toembodiments described on rats subjected to contusive traumatic braininjury and invasive brain injury.

FIGS. 83A and 83B illustrate the effect of a PEMF treatment according toembodiments described on post-operative breast reduction patients.

FIG. 84 illustrates the proportional relationship between levels of1L-1β and force in the Marmarou weight-drop model.

FIG. 85 illustrates the effect of a PEMF treatment according toembodiments described on wound exudate volumes in post-operativepatients under breast reduction surgery.

FIGS. 86A and 86B illustrate PEMF signal configurations according tosome embodiments described.

FIG. 87 illustrates the effect of a PEMF treatment according toembodiments described on inflammation in response to transplants ofdissociated embryonic midbrain neurons.

FIG. 88 illustrates the effect of a PEMF treatment according toembodiments described on microglia in rats subjected to penetratinginjuries.

FIG. 89 illustrates the effect of a PEMF treatment according toembodiments described on dopaminergic neurons.

DETAILED DESCRIPTION Part 1

Induced time-varying currents from PEMF or PRF devices flow in a targetpathway structure such as a molecule, cell, tissue, and organ, and it isthese currents that are a stimulus to which cells and tissues can reactin a physiologically meaningful manner. The electrical properties of atarget pathway structure affect levels and distributions of inducedcurrent. Molecules, cells, tissue, and organs are all in an inducedcurrent pathway such as cells in a gap junction contact. Ion or ligandinteractions at binding sites on macromolecules that may reside on amembrane surface are voltage dependent processes, that iselectrochemical, that can respond to an induced electromagnetic field(“E”). Induced current arrives at these sites via a surrounding ionicmedium. The presence of cells in a current pathway causes an inducedcurrent (“J”) to decay more rapidly with time (“J(t)”). This is due toan added electrical impedance of cells from membrane capacitance andtime constants of binding and other voltage sensitive membrane processessuch as membrane transport.

Equivalent electrical circuit models representing various membrane andcharged interface configurations have been derived. For example, inCalcium (“Ca²⁺”) binding, the change in concentration of bound Ca²⁺ at abinding site due to induced E may be described in a frequency domain byan impedance expression such as:

${Z_{b}(\omega)} = {R_{ion} + \frac{1}{i\; \omega \; C_{ion}}}$

which has the form of a series resistance-capacitance electricalequivalent circuit. Where ω is angular frequency defined as 2πf, where fis frequency, i=−1^(1/2), Z_(b)(ω) is the binding impedance, and R_(ion)and C_(ion) are equivalent binding resistance and capacitance of an ionbinding pathway. The value of the equivalent binding time constant,τ_(ion)=R_(ion)C_(ion), is related to a ion binding rate constant,k_(b), via τ_(ion)=R_(ion)C_(ion)=1/k_(b). Thus, the characteristic timeconstant of this pathway is determined by ion binding kinetics.

Induced E from a PEMF or PRF signal can cause current to flow into anion binding pathway and affect the number of Ca²⁺ ions bound per unittime. An electrical equivalent of this is a change in voltage across theequivalent binding capacitance C_(ion), which is a direct measure of thechange in electrical charge stored by C_(ion). Electrical charge isdirectly proportional to a surface concentration of Ca²⁺ ions in thebinding site, that is storage of charge is equivalent to storage of ionsor other charged species on cell surfaces and junctions. Electricalimpedance measurements, as well as direct kinetic analyses of bindingrate constants, provide values for time constants necessary forconfiguration of a PMF waveform to match a bandpass of target pathwaystructures. This allows for a required range of frequencies for anygiven induced E waveform for optimal coupling to target impedance, suchas bandpass.

Ion binding to regulatory molecules is a frequent EMF target, forexample Ca²⁺ binding to calmodulin (“CaM”). Use of this pathway is basedupon acceleration of wound repair, for example bone repair, thatinvolves modulation of growth factors released in various stages ofrepair. Growth factors such as platelet derived growth factor (“PDGF”),fibroblast growth factor (“FGF”), and epidermal growth factor (“EGF”)are all involved at an appropriate stage of healing. Angiogenesis isalso integral to wound repair and modulated by PMF. All of these factorsare Ca/CaM-dependent.

Utilizing a Ca/CaM pathway a waveform can be configured for whichinduced power is sufficiently above background thermal noise power.Under correct physiological conditions, this waveform can have aphysiologically significant bioeffect.

Application of a Power SNR model to Ca/CaM requires knowledge ofelectrical equivalents of Ca²⁺ binding kinetics at CaM. Within firstorder binding kinetics, changes in concentration of bound Ca²⁺ at CaMbinding sites over time may be characterized in a frequency domain by anequivalent binding time constant, τ_(ion)=R_(ion)C_(ion), where R_(ion)and C_(ion) are equivalent binding resistance and capacitance of the ionbinding pathway. τ_(ion) is related to a ion binding rate constant,k_(b), via τ_(ion)=R_(ion) C_(ion)=1/k_(b). Published values for k_(b)can then be employed in a cell array model to evaluate SNR by comparingvoltage induced by a PRF signal to thermal fluctuations in voltage at aCaM binding site. Employing numerical values for PMF response, such asV_(max)=6.5×10⁻⁷ sec⁻¹, =2.5 μM, K_(D)=30 μM, [Ca²⁺CaM]=K_(D)(+[CaM]),yields k_(b)=665 sec⁻¹ (τ_(ion)=1.5 msec). Such a value for τ_(ion) canbe employed in an electrical equivalent circuit for ion binding whilepower SNR analysis can be performed for any waveform structure.

According to an embodiment of the present invention a mathematical modelcan be configured to assimilate that thermal noise is present in allvoltage dependent processes and represents a minimum thresholdrequirement to establish adequate SNR. Power spectral density, S_(n)(ω),of thermal noise can be expressed as:

S _(n)(ω)=4kT Re[Z _(M)(x,ω)]

where Z_(M)(x, ω) is electrical impedance of a target pathway structure,x is a dimension of a target pathway structure and Re denotes a realpart of impedance of a target pathway structure. Z_(M)(x, ω) can beexpressed as:

${Z_{M}( {x,\omega} )} = {\lbrack \frac{R_{e} + R_{i} + R_{g}}{y} \rbrack \tanh \; ( {y\; x} )}$

This equation clearly shows that electrical impedance of the targetpathway structure, and contributions from extracellular fluid resistance(“R_(e)”), intracellular fluid resistance (“R_(i)”) and intermembraneresistance (“R_(g)”) which are electrically connected to a targetpathway structure, all contribute to noise filtering.

A typical approach to evaluation of SNR uses a single value of a rootmean square (RMS) noise voltage. This is calculated by taking a squareroot of an integration of S_(n)(ω)=4 kT Re [Z_(M)(x, ω)] over allfrequencies relevant to either complete membrane response, or tobandwidth of a target pathway structure. SNR can be expressed by aratio:

${SNR} = \frac{{V_{M}(\omega)}}{RMS}$

where |V_(M)(ω)| is maximum amplitude of voltage at each frequency asdelivered by a chosen waveform to the target pathway structure.

Referring to FIG. 1, wherein FIG. 1 is a flow diagram of a method fordelivering electromagnetic signals to target pathway structures such asmolecules, cells, tissue and organs of plants, animals, and humans fortherapeutic and prophylactic purposes according to an embodiment of thepresent invention. A mathematical model having at least one waveformparameter is applied to configure at least one waveform to be coupled toa target pathway structure such as a molecule, cell, tissue, and organ(Step 101). The configured waveform satisfies a SNR or Power SNR modelso that for a given and known target pathway structure it is possible tochoose at least one waveform parameter so that a waveform is detectablein the target pathway structure above its background activity (Step 102)such as baseline thermal fluctuations in voltage and electricalimpedance at a target pathway structure that depend upon a state of acell and tissue, that is whether the state is at least one of resting,growing, replacing, and responding to injury. A preferred embodiment ofa generated electromagnetic signal is comprised of a burst of arbitrarywaveforms having at least one waveform parameter that includes aplurality of frequency components ranging from about 0.01 Hz to about100 MHz wherein the plurality of frequency components satisfies a PowerSNR model (Step 102). A repetitive electromagnetic signal can begenerated for example inductively or capacitively, from said configuredat least one waveform (Step 103). The electromagnetic signal is coupledto a target pathway structure such as a molecule, cell, tissue, andorgan by output of a coupling device such as an electrode or aninductor, placed in close proximity to the target pathway structure(Step 104). The coupling enhances a stimulus to which cells and tissuesreact in a physiologically meaningful manner.

FIG. 2 illustrates a preferred embodiment of an apparatus according tothe present invention. A miniature control circuit 201 is coupled to anend of at least one connector 202 such as wire. The opposite end of theat least one connector is coupled to a generating device such as a pairof electrical coils 203. The miniature control circuit 201 isconstructed in a manner that applies a mathematical model that is usedto configure waveforms. The configured waveforms have to satisfy a SNRor Power SNR model so that for a given and known target pathwaystructure, it is possible to choose waveform parameters that satisfy SNRor Power SNR so that a waveform is detectable in the target pathwaystructure above its background activity. A preferred embodimentaccording to the present invention applies a mathematical model toinduce a time-varying magnetic field and a time-varying electric fieldin a target pathway structure such as a molecule, cell, tissue, andorgan, comprising about 10 to about 100 msec bursts of about 1 to about100 microsecond rectangular pulses repeating at about 0.1 to about 10pulses per second. Peak amplitude of the induced electric field isbetween about 1 uV/cm and about 100 mV/cm, varied according to amodified 1/f function where f=frequency. A waveform configured using apreferred embodiment according to the present invention may be appliedto a target pathway structure such as a molecule, cell, tissue, andorgan for a preferred total exposure time of under 1 minute to 240minutes daily. However other exposure times can be used. Waveformsconfigured by the miniature control circuit 201 are directed to agenerating device 203 such as electrical coils via connector 202. Thegenerating device 203 delivers a pulsing magnetic field configuredaccording to a mathematical model, that can be used to provide treatmentto a target pathway structure such as knee joint 204. The miniaturecontrol circuit applies a pulsing magnetic field for a prescribed timeand can automatically repeat applying the pulsing magnetic field for asmany applications as are needed in a given time period, for example 10times a day. A preferred embodiment according to the present inventioncan be positioned to treat the knee joint 204 by a positioning device.The positioning device can be portable such as an anatomical support,and is further described below with reference to FIG. 6. Coupling apulsing magnetic field to a target pathway structure such as a molecule,cell, tissue, and organ, therapeutically and prophylactically reducesinflammation thereby reducing pain and promotes healing. When electricalcoils are used as the generating device 203, the electrical coils can bepowered with a time varying magnetic field that induces a time varyingelectric field in a target pathway structure according to Faraday's law.An electromagnetic signal generated by the generating device 203 canalso be applied using electrochemical coupling, wherein electrodes arein direct contact with skin or another outer electrically conductiveboundary of a target pathway structure. Yet in another embodimentaccording to the present invention, the electromagnetic signal generatedby the generating device 203 can also be applied using electrostaticcoupling wherein an air gap exists between a generating device 203 suchas an electrode and a target pathway structure such as a molecule, cell,tissue, and organ. An advantage of the preferred embodiment according tothe present invention is that its ultra lightweight coils andminiaturized circuitry allow for use with common physical therapytreatment modalities and at any body location for which pain relief andhealing is desired. An advantageous result of application of thepreferred embodiment according to the present invention is that a livingorganism's wellbeing can be maintained and enhanced.

FIG. 3 depicts a block diagram of a preferred embodiment according tothe present invention of a miniature control circuit 300. The miniaturecontrol circuit 300 produces waveforms that drive a generating devicesuch as wire coils described above in FIG. 2. The miniature controlcircuit can be activated by any activation means such as an on/offswitch. The miniature control circuit 300 has a power source such as alithium battery 301. A preferred embodiment of the power source has anoutput voltage of 3.3 V but other voltages can be used. In anotherembodiment according to the present invention the power source can be anexternal power source such as an electric current outlet such as anAC/DC outlet, coupled to the present invention for example by a plug andwire. A switching power supply 302 controls voltage to amicro-controller 303. A preferred embodiment of the micro-controller 303uses an 8 bit 4 MHz micro-controller 303 but other bit MHz combinationmicro-controllers may be used. The switching power supply 302 alsodelivers current to storage capacitors 304. A preferred embodiment ofthe present invention uses storage capacitors having a 220 uF output butother outputs can be used. The storage capacitors 304 allow highfrequency pulses to be delivered to a coupling device such as inductors(Not Shown). The micro-controller 303 also controls a pulse shaper 305and a pulse phase timing control 306. The pulse shaper 305 and pulsephase timing control 306 determine pulse shape, burst width, burstenvelope shape, and burst repetition rate. An integral waveformgenerator, such as a sine wave or arbitrary number generator can also beincorporated to provide specific waveforms. A voltage level conversionsub-circuit 308 controls an induced field delivered to a target pathwaystructure. A switching Hexfet 308 allows pulses of randomized amplitudeto be delivered to output 309 that routes a waveform to at least onecoupling device such as an inductor. The micro-controller 303 can alsocontrol total exposure time of a single treatment of a target pathwaystructure such as a molecule, cell, tissue, and organ. The miniaturecontrol circuit 300 can be constructed to apply a pulsing magnetic fieldfor a prescribed time and to automatically repeat applying the pulsingmagnetic field for as many applications as are needed in a given timeperiod, for example 10 times a day. A preferred embodiment according tothe present invention uses treatments times of about 10 minutes to about30 minutes.

Referring to FIGS. 4A and 4B a preferred embodiment according to thepresent invention of a coupling device 400 such as an inductor is shown.The coupling device 400 can be an electric coil 401 wound withmultistrand flexible magnetic wire 402. The multistrand flexiblemagnetic wire 402 enables the electric coil 401 to conform to specificanatomical configurations such as a limb or joint of a human or animal.A preferred embodiment of the electric coil 401 comprises about 10 toabout 50 turns of about 0.01 mm to about 0.1 mm diameter multistrandmagnet wire wound on an initially circular form having an outer diameterbetween about 2.5 cm and about 50 cm but other numbers of turns and wirediameters can be used. A preferred embodiment of the electric coil 401can be encased with a non-toxic PVC mould 403 but other non-toxic mouldscan also be used.

Referring to FIG. 5 an embodiment according to the present invention ofa waveform 500 is illustrated. A pulse 501 is repeated within a burst502 that has a finite duration 503. The duration 503 is such that a dutycycle which can be defined as a ratio of burst duration to signal periodis between about 1 to about 10⁻⁵. A preferred embodiment according tothe present invention utilizes pseudo rectangular 10 microsecond pulsesfor pulse 501 applied in a burst 502 for about 10 to about 50 msechaving a modified 1/f amplitude envelope 504 and with a finite duration503 corresponding to a burst period of between about 0.1 and about 10seconds.

FIG. 6 illustrates a preferred embodiment according to the presentinvention of a positioning device such as a wrist support. A positioningdevice 600 such as a wrist support 601 is worn on a human wrist 602. Thepositioning device can be constructed to be portable, can be constructedto be disposable, and can be constructed to be implantable. Thepositioning device can be used in combination with the present inventionin a plurality of ways, for example incorporating the present inventioninto the positioning device for example by stitching, affixing thepresent invention onto the positioning device for example by Velcro®,and holding the present invention in place by constructing thepositioning device to be elastic.

In another embodiment according to the present invention, the presentinvention can be constructed as a stand-alone device of any size with orwithout a positioning device, to be used anywhere for example at home,at a clinic, at a treatment center, and outdoors. The wrist support 601can be made with any anatomical and support material, such as neoprene.Coils 603 are integrated into the wrist support 601 such that a signalconfigured according to the present invention, for example the waveformdepicted in FIG. 5, is applied from a dorsal portion that is the top ofthe wrist to a plantar portion that is the bottom of the wrist.Micro-circuitry 604 is attached to the exterior of the wrist support 601using a fastening device such as Velcro® (Not Shown). Themicro-circuitry is coupled to one end of at least one connecting devicesuch as a flexible wire 605. The other end of the at least oneconnecting device is coupled to the coils 603. Other embodimentsaccording to the present invention of the positioning device includeknee, elbow, lower back, shoulder, other anatomical wraps, and apparelsuch as garments, fashion accessories, and footware.

Example 1

The Power SNR approach for PMF signal configuration has been testedexperimentally on calcium dependent myosin phosphorylation in a standardenzyme assay. The cell-free reaction mixture was chosen forphosphorylation rate to be linear in time for several minutes, and forsub-saturation Ca²⁺ concentration. This opens the biological window forCa²⁺/CaM to be EMF-sensitive. This system is not responsive to PMF atlevels utilized in this study if Ca²⁺ is at saturation levels withrespect to CaM, and reaction is not slowed to a minute time range.Experiments were performed using myosin light chain (“MLC”) and myosinlight chain kinase (“MLCK”) isolated from turkey gizzard. A reactionmixture consisted of a basic solution containing 40 mM Hepes buffer, pH7.0; 0.5 mM magnesium acetate; 1 mg/ml bovine serum albumin, 0.1% (w/v)Tween 80; and 1 mM EGTA12. Free Ca²⁺ was varied in the 1-7 μM range.Once Ca²⁺ buffering was established, freshly prepared 70 nM CaM, 160 nMMLC and 2 nM MLCK were added to the basic solution to form a finalreaction mixture. The low MLC/MLCK ratio allowed linear time behavior inthe minute time range. This provided reproducible enzyme activities andminimized pipetting time errors.

The reaction mixture was freshly prepared daily for each series ofexperiments and was aliquoted in 100 μL portions into 1.5 ml Eppendorftubes. All Eppendorf tubes containing reaction mixture were kept at 0°C. then transferred to a specially designed water bath maintained at37±0.1° C. by constant perfusion of water prewarmed by passage through aFisher Scientific model 900 heat exchanger. Temperature was monitoredwith a thermistor probe such as a Cole-Parmer model 8110-20, immersed inone Eppendorf tube during all experiments. Reaction was initiated with2.5 μM 32P ATP, and was stopped with Laemmli Sample Buffer solutioncontaining 30 μM EDTA. A minimum of five blank samples were counted ineach experiment. Blanks comprised a total assay mixture minus one of theactive components Ca²⁺, CaM, MLC or MLCK. Experiments for which blankcounts were higher than 300 cpm were rejected. Phosphorylation wasallowed to proceed for 5 min and was evaluated by counting 32Pincorporated in MLC using a TM Analytic model 5303 Mark V liquidscintillation counter.

The signal comprised repetitive bursts of a high frequency waveform.Amplitude was maintained constant at 0.2 G and repetition rate was 1burst/sec for all exposures. Burst duration varied from 65 μsec to 1000μsec based upon projections of Power SNR analysis which showed thatoptimal Power SNR would be achieved as burst duration approached 500μsec. The results are shown in FIG. 7 wherein burst width 701 in μsec isplotted on the x-axis and Myosin Phosphorylation 702 as treated/sham isplotted on the y-axis. It can be seen that the PMF effect on Ca²⁺binding to CaM approaches its maximum at approximately 500 μsec, just asillustrated by the Power SNR model.

These results confirm that a PMF signal, configured according to anembodiment of the present invention, would maximally increase myosinphosphorylation for burst durations sufficient to achieve optimal PowerSNR for a given magnetic field amplitude.

Example 2

According to an embodiment of the present invention use of a Power SNRmodel was further verified in an in vivo wound repair model. A rat woundmodel has been well characterized both biomechanically andbiochemically, and was used in this study. Healthy, young adult maleSprague Dawley rats weighing more than 300 grams were utilized.

The animals were anesthetized with an intraperitoneal dose of Ketamine75 mg/kg and Medetomidine 0.5 mg/kg. After adequate anesthesia had beenachieved, the dorsum was shaved, prepped with a dilute betadine/alcoholsolution, and draped using sterile technique. Using a #10 scalpel, an8-cm linear incision was performed through the skin down to the fasciaon the dorsum of each rat. The wound edges were bluntly dissected tobreak any remaining dermal fibers, leaving an open wound approximately 4cm in diameter. Hemostasis was obtained with applied pressure to avoidany damage to the skin edges. The skin edges were then closed with a 4-0Ethilon running suture. Post-operatively, the animals receivedBuprenorphine 0.1-0.5 mg/kg, intraperitoneal. They were placed inindividual cages and received food and water ad libitum.

PMF exposure comprised two pulsed radio frequency waveforms. The firstwas a standard clinical PRF signal comprising a 65 μsec burst of 27.12MHz sinusoidal waves at 1 Gauss amplitude and repeating at 600bursts/sec. The second was a PRF signal reconfigured according to anembodiment of the present invention. For this signal burst duration wasincreased to 2000 μsec and the amplitude and repetition rate werereduced to 0.2 G and 5 bursts/sec respectively. PRF was applied for 30minutes twice daily.

Tensile strength was performed immediately after wound excision. Two 1cm width strips of skin were transected perpendicular to the scar fromeach sample and used to measure the tensile strength in kg/mm². Thestrips were excised from the same area in each rat to assure consistencyof measurement. The strips were then mounted on a tensiometer. Thestrips were loaded at 10 mm/min and the maximum force generated beforethe wound pulled apart was recorded. The final tensile strength forcomparison was determined by taking the average of the maximum load inkilograms per mm² of the two strips from the same wound.

The results showed average tensile strength for the 65 μsec 1 Gauss PRFsignal was 19.3±4.3 kg/mm² for the exposed group versus 13.0±3.5 kg/mm²for the control group (p<0.01), which is a 48% increase. In contrast,the average tensile strength for the 2000 μsec 0.2 Gauss PRF signal,configured according to an embodiment of the present invention using aPower SNR model was 21.2±5.6 kg/mm² for the treated group versus13.7±4.1 kg/mm² (p<0.01) for the control group, which is a 54% increase.The results for the two signals were not significantly different fromeach other.

These results demonstrate that an embodiment of the present inventionallowed a new PRF signal to be configured that could be produced withsignificantly lower power. The PRF signal configured according to anembodiment of the present invention, accelerated wound repair in the ratmodel in a low power manner versus that for a clinical PRF signal whichaccelerated wound repair but required more than two orders of magnitudemore power to produce.

Example 3

In this example Jurkat cells react to PMF stimulation of a T-cellreceptor with cell cycle arrest and thus behave like normalT-lymphocytes stimulated by antigens at the T-cell receptor such asanti-CD3. For example in bone healing, results have shown both 60 Hz andPEMF fields decrease DNA synthesis of Jurkat cells, as is expected sincePMF interacts with the T-cell receptor in the absence of a costimulatorysignal. This is consistent with an anti-inflammatory response, as hasbeen observed in clinical applications of PMF stimuli. The PEMF signalis more effective. A dosimetry analysis performed according to anembodiment of the present invention demonstrates why both signals areeffective and why PEMF signals have a greater effect than 60 Hz signalson Jurkat cells in the most EMF-sensitive growth stage.

Comparison of dosimetry from the two signals employed involvesevaluation of the ratio of the Power spectrum of the thermal noisevoltage that is Power SNR, to that of the induced voltage at theEMF-sensitive target pathway structure. The target pathway structureused is ion binding at receptor sites on Jurkat cells suspended in 2 mmof culture medium. The average peak electric field at the binding sitefrom a PEMF signal comprising 5 msec burst of 200 μsec pulses repeatingat 15/sec, was 1 mV/cm, while for a 60 Hz signal it was 50 μV/cm.

FIG. 8 is a graph of results wherein Induced Field Frequency 801 in Hzis plotted on the x-axis and Power SNR 802 is plotted on the y-axis.FIG. 8 illustrates that both signals have sufficient Power spectrum thatis Power SNR≈1, to be detected within a frequency range of bindingkinetics. However, maximum Power SNR for the PEMF signal issignificantly higher than that for the 60 Hz signal. This is because aPEMF signal has many frequency components falling within the bandpass ofthe binding pathway. The single frequency component of a 60 Hz signallies at the mid-point of the bandpass of the target pathway. The PowerSNR calculation that was used in this example is dependant upon τ_(ion)which is obtained from the rate constant for ion binding. Had thiscalculation been performed a priori it would have concluded that bothsignals satisfied basic detectability requirements and could modulate anEMF-sensitive ion binding pathway at the start of a regulatory cascadefor DNA synthesis in these cells. The previous examples illustrated thatutilizing the rate constant for Ca/CaM binding could lead to successfulprojections for bioeffective EMF signals in a variety of systems.

Having described embodiments for an apparatus and a method fordelivering electromagnetic treatment to human, animal and plantmolecules, cells, tissue and organs, it is noted that modifications andvariations can be made by persons skilled in the art in light of theabove teachings. It is therefore to be understood that changes may bemade in the particular embodiments of the invention disclosed which arewithin the scope and spirit of the invention as defined by the appendedclaims.

Part 2

Induced time-varying currents from PEMF or PRF devices flow in a targetpathway structure such as a molecule, cell, tissue, and organ, and it isthese currents that are a stimulus to which cells and tissues can reactin a physiologically meaningful manner. The electrical properties of atarget pathway structure affect levels and distributions of inducedcurrent. Molecules, cells, tissue, and organs are all in an inducedcurrent pathway such as cells in a gap junction contact. Ion or ligandinteractions at binding sites on macromolecules that may reside on amembrane surface are voltage dependent processes, that iselectrochemical, that can respond to an induced electromagnetic field(“E”). Induced current arrives at these sites via a surrounding ionicmedium. The presence of cells in a current pathway causes an inducedcurrent (“J”) to decay more rapidly with time (“J(t)”). This is due toan added electrical impedance of cells from membrane capacitance andtime constants of binding and other voltage sensitive membrane processessuch as membrane transport.

Equivalent electrical circuit models representing various membrane andcharged interface configurations have been derived. For example, inCalcium (“Ca²⁺”) binding, the change in concentration of bound Ca²⁺ at abinding site due to induced E may be described in a frequency domain byan impedance expression such as:

${Z_{b}(\omega)} = {R_{ion} + \frac{1}{i\; \omega \; C_{ion}}}$

which has the form of a series resistance-capacitance electricalequivalent circuit. Where ω is angular frequency defined as 2πf, where fis frequency, i=−1^(1/2), Z_(b)(ω) is the binding impedance, and R_(ion)and C_(ion) are equivalent binding resistance and capacitance of an ionbinding pathway. The value of the equivalent binding time constant,τ_(ion)=R_(ion)C_(ion), is related to a ion binding rate constant,k_(b), via τ_(ion)=R_(ion)C_(ion)=1/k_(b). Thus, the characteristic timeconstant of this pathway is determined by ion binding kinetics.

Induced E from a PEMF or PRF signal can cause current to flow into anion binding pathway and affect the number of Ca²⁺ ions bound per unittime. An electrical equivalent of this is a change in voltage across theequivalent binding capacitance C_(ion), which is a direct measure of thechange in electrical charge stored by C_(ion). Electrical charge isdirectly proportional to a surface concentration of Ca²⁺ ions in thebinding site, that is storage of charge is equivalent to storage of ionsor other charged species on cell surfaces and junctions. Electricalimpedance measurements, as well as direct kinetic analyses of bindingrate constants, provide values for time constants necessary forconfiguration of a PMF waveform to match a bandpass of target pathwaystructures. This allows for a required range of frequencies for anygiven induced E waveform for optimal coupling to target impedance, suchas bandpass.

Ion binding to regulatory molecules is a frequent EMF target, forexample Ca²⁺ binding to calmodulin (“CaM”). Use of this pathway is basedupon acceleration of wound repair, for example bone repair, thatinvolves modulation of growth factors released in various stages ofrepair. Growth factors such as platelet derived growth factor (“PDGF”),fibroblast growth factor (“FGF”), and epidermal growth factor (“EGF”)are all involved at an appropriate stage of healing. Angiogenesis andneovascularization are also integral to wound repair and can bemodulated by PMF. All of these factors are Ca/CaM-dependent.

Utilizing a Ca/CaM pathway a waveform can be configured for whichinduced power is sufficiently above background thermal noise power.Under correct physiological conditions, this waveform can have aphysiologically significant bioeffect.

Application of a Power SNR model to Ca/CaM requires knowledge ofelectrical equivalents of Ca²⁺ binding kinetics at CaM. Within firstorder binding kinetics, changes in concentration of bound Ca²⁺ at CaMbinding sites over time may be characterized in a frequency domain by anequivalent binding time constant, τ_(ion)=R_(ion) C_(ion), where R_(ion)and C_(ion) are equivalent binding resistance and capacitance of the ionbinding pathway. τ_(ion) is related to a ion binding rate constant,k_(b), via τ_(ion)=R_(ion) C_(ion)=1/k_(b). Published values for k_(b)can then be employed in a cell array model to evaluate SNR by comparingvoltage induced by a PRF signal to thermal fluctuations in voltage at aCaM binding site. Employing numerical values for PMF response, such asV_(max)=6.5×10⁻⁷ sec⁻¹, =2.50/1, K_(D)=30 μM, [Ca²⁺CaM]=K_(D)(+[CaM]),yields k_(b)=665 sec⁻¹ (τ_(ion)=1.5 msec). Such a value for τ_(ion) canbe employed in an electrical equivalent circuit for ion binding whilepower SNR analysis can be performed for any waveform structure.

According to an embodiment of the present invention a mathematical modelcan be configured to assimilate that thermal noise is present in allvoltage dependent processes and represents a minimum thresholdrequirement to establish adequate SNR. Power spectral density, S_(n)(ω),of thermal noise can be expressed as:

S _(n)(ω)=4kT Re[Z _(M)(x,ω)]

where Z_(M)(x,ω) is electrical impedance of a target pathway structure,x is a dimension of a target pathway structure and Re denotes a realpart of impedance of a target pathway structure. Z_(M)(x,ω) can beexpressed as:

${Z_{M}( {x,\omega} )} = {\lbrack \frac{R_{e} + R_{i} + R_{g}}{\gamma} \rbrack {\tanh ( {\gamma \; x} )}}$

This equation clearly shows that electrical impedance of the targetpathway structure, and contributions from extracellular fluid resistance(“R_(e)”), intracellular fluid resistance (“R_(i)”) and intermembraneresistance (“R_(g)”) which are electrically connected to a targetpathway structure, all contribute to noise filtering.

A typical approach to evaluation of SNR uses a single value of a rootmean square (RMS) noise voltage. This is calculated by taking a squareroot of an integration of

S_(n)(ω)=4 kT Re[Z_(M)(x, ω)] over all frequencies relevant to eithercomplete membrane response, or to bandwidth of a target pathwaystructure. SNR can be expressed by a ratio:

${SNR} = \frac{{V_{M}(\omega)}}{RMS}$

where |V_(M)(ω)| is maximum amplitude of voltage at each frequency asdelivered by a chosen waveform to the target pathway structure.

An embodiment according to the present invention comprises a pulse burstenvelope having a high spectral density, so that the effect of therapyupon the relevant dielectric pathways, such as, cellular membranereceptors, ion binding to cellular enzymes and general transmembranepotential changes, is enhanced. Accordingly by increasing a number offrequency components transmitted to relevant cellular pathways, a largerange of biophysical phenomena, such as modulating growth factor andcytokine release and ion binding at regulatory molecules, applicable toknown healing mechanisms is accessible. According to an embodiment ofthe present invention applying a random, or other high spectral densityenvelope, to a pulse burst envelope of mono- or bi-polar rectangular orsinusoidal pulses inducing peak electric fields between about 10⁻⁶ andabout 100 V/cm, produces a greater effect on biological healingprocesses applicable to both soft and hard tissues.

According to yet another embodiment of the present invention by applyinga high spectral density voltage envelope as a modulating or pulse-burstdefining parameter, power requirements for such amplitude modulatedpulse bursts can be significantly lower than that of an unmodulatedpulse burst containing pulses within a similar frequency range. This isdue to a substantial reduction in duty cycle within repetitive bursttrains brought about by imposition of an irregular, and preferablyrandom, amplitude onto what would otherwise be a substantially uniformpulse burst envelope. Accordingly, the dual advantages, of enhancedtransmitted dosimetry to the relevant dielectric pathways and ofdecreased power requirement are achieved.

Referring to FIG. 9, wherein FIG. 9 is a flow diagram of a method ofusing an inductive apparatus to deliver electromagnetic signals totarget pathway structures such as such as molecules, cells, tissues, andorgans of plants, animals, and humans for therapeutic and prophylacticpurposes according to an embodiment of the present invention. Alightweight inductive apparatus is integrated into at least onetherapeutic device that will be used for treatment, however theinductive apparatus can also be attached to at least one therapeuticdevice (Step 9101). Miniaturized circuitry containing logic for amathematical model having at least one waveform parameter used toconfigure at least one waveform to be coupled to a target pathwaystructure such as molecules, cells, tissues, and organs, is attached tothe coil by at least one wire (Step 9102). However, the attachment canalso be wireless. The configured waveform satisfies a SNR or Power SNRmodel so that for a given and known target pathway structure it ispossible to choose at least one waveform parameter so that a waveform isdetectable in the target pathway structure above its background activity(Step 9103) such as baseline thermal fluctuations in voltage andelectrical impedance at a target pathway structure that depend upon astate of a cell and tissue, that is whether the state is at least one ofresting, growing, replacing, and responding to injury. A preferredembodiment of a generated electromagnetic signal is comprised of a burstof arbitrary waveforms having at least one waveform parameter thatincludes a plurality of frequency components ranging from about 0.01 Hzto about 100 MHz wherein the plurality of frequency components satisfiesa Power SNR model (Step 9104). A repetitive electromagnetic signal canbe generated for example inductively, from said configured at least onewaveform (Step 9105). The repetitive electromagnetic signal can also begenerated conductively. The electromagnetic signal is coupled to atarget pathway structure such as molecules, cells, tissues, and organsby output of the inductive apparatus integrated into the support (Step9106).

FIG. 10 illustrates a preferred embodiment of an apparatus according tothe present invention. A miniature control circuit 10201 is coupled toan end of at least one connector 10202 such as wire. The opposite end ofthe at least one connector is coupled to a generating device such as apair of electrical coils 10203. The generating device is constructed tohave electrical properties that optimize generation of electromagneticsignals from waveforms configured to satisfy at least one of a SNRmodel, a Power SNR model, and any other mathematical model used forwaveform configuration. The miniature control circuit 10201 isconstructed in a manner that applies a mathematical model that is usedto configure waveforms. The configured waveforms have to satisfy a SNRor Power SNR model so that for a given and known target pathwaystructure, it is possible to choose waveform parameters that satisfy SNRor Power SNR so that a waveform is detectable in the target pathwaystructure above its background activity. A preferred embodimentaccording to the present invention applies a mathematical model toinduce a time-varying magnetic field and a time-varying electric fieldin a target pathway structure such as molecules, cells, tissues, andorgans, comprising about 10 to about 100 msec bursts of about 1 to about100 microsecond rectangular pulses repeating at about 0.1 to about 10pulses per second. Peak amplitude of the induced electric field isbetween about 1 uV/cm and about 100 mV/cm, varied according to amodified 1/f function where f=frequency. A waveform configured using apreferred embodiment according to the present invention may be appliedto a target pathway structure such as molecules, cells, tissues, andorgans for a preferred total exposure time of under 1 minute to 240minutes daily. However other exposure times can be used. Waveformsconfigured by the miniature control circuit 10201 are directed to agenerating device 10203 such as electrical coils via connector 10202.The generating device 10203 delivers a pulsing magnetic field configuredaccording to a mathematical model, that can be used to provide treatmentto a target pathway structure such as a heart in a chest 10204. Theminiature control circuit applies a pulsing magnetic field for aprescribed time and can automatically repeat applying the pulsingmagnetic field for as many applications as are needed in a given timeperiod, for example 10 times a day. A preferred embodiment according tothe present invention can be positioned to treat the heart in a chest10204 by a positioning device. Coupling a pulsing magnetic field to aangiogenesis and neovascularization target pathway structure such asions and ligands, therapeutically and prophylactically reducesinflammation thereby reducing pain and promotes healing. When electricalcoils are used as the generating device 10203, the electrical coils canbe powered with a time varying magnetic field that induces a timevarying electric field in a target pathway structure according toFaraday's law. An electromagnetic signal generated by the generatingdevice 10203 can also be applied using electrochemical coupling, whereinelectrodes are in direct contact with skin or another outer electricallyconductive boundary of a target pathway structure. Yet in anotherembodiment according to the present invention, the electromagneticsignal generated by the generating device 10203 can also be appliedusing electrostatic coupling wherein an air gap exists between agenerating device 10203 such as an electrode and a target pathwaystructure such as molecules, cells, tissues, and organs. An advantage ofthe preferred embodiment according to the present invention is that itsultra lightweight coils and miniaturized circuitry allow for use withcommon physical therapy treatment modalities and at any body locationfor which pain relief and healing is desired. An advantageous result ofapplication of the preferred embodiment according to the presentinvention is that a living organism's angiogenesis andneovascularization can be maintained and enhanced.

FIG. 11 depicts a block diagram of a preferred embodiment according tothe present invention of a miniature control circuit 11300. Theminiature control circuit 11300 produces waveforms that drive agenerating device such as wire coils described above in FIG. 10. Theminiature control circuit can be activated by any activation means suchas an on/off switch. The miniature control circuit 11300 has a powersource such as a lithium battery 11301. A preferred embodiment of thepower source has an output voltage of 3.3 V but other voltages can beused. In another embodiment according to the present invention the powersource can be an external power source such as an electric currentoutlet such as an AC/DC outlet, coupled to the present invention forexample by a plug and wire. A switching power supply 11302 controlsvoltage to a micro-controller 11303. A preferred embodiment of themicro-controller 11303 uses an 8 bit 4 MHz micro-controller 11303 butother bit MHz combination micro-controllers may be used. The switchingpower supply 11302 also delivers current to storage capacitors 11304. Apreferred embodiment of the present invention uses storage capacitorshaving a 220 uF output but other outputs can be used. The storagecapacitors 11304 allow high frequency pulses to be delivered to acoupling device such as inductors (Not Shown). The micro-controller11303 also controls a pulse shaper 11305 and a pulse phase timingcontrol 11306. The pulse shaper 11305 and pulse phase timing control11306 determine pulse shape, burst width, burst envelope shape, andburst repetition rate. An integral waveform generator, such as a sinewave or arbitrary number generator can also be incorporated to providespecific waveforms. A voltage level conversion sub-circuit 11308controls an induced field delivered to a target pathway structure. Aswitching Hexfet 11308 allows pulses of randomized amplitude to bedelivered to output 11309 that routes a waveform to at least onecoupling device such as an inductor. The micro-controller 11303 can alsocontrol total exposure time of a single treatment of a target pathwaystructure such as a molecule, cell, tissue, and organ. The miniaturecontrol circuit 11300 can be constructed to apply a pulsing magneticfield for a prescribed time and to automatically repeat applying thepulsing magnetic field for as many applications as are needed in a giventime period, for example 10 times a day. A preferred embodimentaccording to the present invention uses treatments times of about 10minutes to about 30 minutes.

Referring to FIG. 12 an embodiment according to the present invention ofan electromagnetic treatment inductive apparatus integrated into hip,thigh, and lower back support garment 12400 is illustrated. Severallightweight flexible coils 12401 are integrated into the supportgarment. The lightweight flexible coils can be constructed from fineflexible conductive wire, conductive thread, and any other flexibleconductive material. The flexible coils are connected to at least oneend of at least one wire 12402. However the flexible coils can also beconfigured to be directly connected to circuitry 12403 or wireless.Lightweight miniaturized circuitry 12403 that configures waveformsaccording to an embodiment of the present invention, is attached to atleast one other end of said at least on wire. When activated thelightweight miniaturized circuitry 12403 configures waveforms that aredirected to the flexible coils (12401) to create PEMF signals that arecoupled to a target pathway structure.

Referring to FIG. 13 an embodiment according to the present invention ofan electromagnetic treatment inductive apparatus integrated into a headand face support garment 13500 is illustrated. Several lightweightflexible coils 13501 are integrated into the support garment. Thelightweight flexible coils can be constructed from fine flexibleconductive wire, conductive thread, and any other flexible conductivematerial. The flexible coils are connected to at least one end of atleast one wire 13502. However, the flexible coils can also be configuredto be directly connected to circuitry 13503 or wireless. Lightweightminiaturized circuitry 13503 that configures waveforms, according to anembodiment of the present invention, is attached to at least one otherend of said at least on wire. When activated the lightweightminiaturized circuitry 503 configures waveforms that are directed to theflexible coils (13501) to create PEMF signals that are coupled to atarget pathway structure.

Referring to FIG. 14 an embodiment according to the present invention ofan electromagnetic treatment inductive apparatus integrated intosurgical dressing applied to a human forearm 14600 is illustrated.Several lightweight flexible coils 14601 are integrated into thedressing. The lightweight flexible coils can be constructed from fineflexible conductive wire, conductive thread, and any other flexibleconductive material. The flexible coils are connected to at least oneend of at least one wire 14602. However, the flexible coils can also beconfigured to be directly connected to circuitry 14603 or wireless.Lightweight miniaturized circuitry 14603 that configures waveformsaccording to an embodiment of the present invention, is attached to atleast one other end of said at least one wire. When activated thelightweight miniaturized circuitry 14603 configures waveforms that aredirected to the flexible coils (14601) to create PEMF signals that arecoupled to a target pathway structure.

Referring to FIG. 15 an embodiment according to the present invention ofan electromagnetic treatment inductive apparatus integrated into amattress pad 15700 is illustrated. Several lightweight flexible coils15701 are integrated into the mattress pad. The lightweight flexiblecoils can be constructed from fine flexible conductive wire, conductivethread, and any other flexible conductive material. The flexible coilsare connected to at least one end of at least one wire 15702. However,the flexible coils can also be configured to be directly connected tocircuitry 15703 or wireless. Lightweight miniaturized circuitry 15703that configures waveforms according to an embodiment of the presentinvention, is attached to at least one other end of said at least onwire. When activated the lightweight miniaturized circuitry 15703configures waveforms that are directed to the flexible coils (15701) tocreate PEMF signals that are coupled to a target pathway structure.

Referring to FIGS. 16A and 16B an embodiment according to the presentinvention of an electromagnetic treatment inductive apparatus integratedinto a sock 16801 and a shoe 16802 are illustrated. Several lightweightflexible coils 16803 are integrated into the dressing. The lightweightflexible coils can be constructed from fine flexible conductive wire,conductive thread, and any other flexible conductive material. Theflexible coils are connected to at least one end of at least one wire16804. However, the flexible coils can also be configured to be directlyconnected to circuitry 16805 or wireless. Lightweight miniaturizedcircuitry 16805 that configures waveforms according to an embodiment ofthe present invention, is attached to at least one other end of said atleast on wire. When activated the lightweight miniaturized circuitry16805 configures waveforms that are directed to the flexible coils(16806) to create PEMF signals that are coupled to a target pathwaystructure.

Referring to FIG. 17 an embodiment according to the present invention ofan electromagnetic treatment inductive apparatus integrated into atherapeutic bed 17900 is illustrated. Several lightweight flexible coils17901 are integrated into the bed. The lightweight flexible coils can beconstructed from fine flexible conductive wire, conductive thread, andany other flexible conductive material. The flexible coils are connectedto at least one end of at least one wire 17902. However, the flexiblecoils can also be configured to be directly connected to circuitry 17903or wireless. Lightweight miniaturized circuitry 17903 that configureswaveforms according to an embodiment of the present invention, isattached to at least one other end of said at least on wire. Whenactivated the lightweight miniaturized circuitry 17903 configureswaveforms that are directed to the flexible coils (17901) to create PEMFsignals that are coupled to a target pathway structure.

Referring to FIG. 18 an embodiment according to the present invention ofan electromagnetic treatment inductive apparatus integrated into a chestgarment 181000, such as a bra is illustrated. Several lightweightflexible coils 181001 are integrated into a bra. The lightweightflexible coils can be constructed from fine flexible conductive wire,conductive thread, and any other flexible conductive material. Theflexible coils are connected to at least one end of at least one wire181002. However, the flexible coils can also be configured to bedirectly connected to circuitry 181003 or wireless. Lightweightminiaturized circuitry 181003 that configures waveforms according to anembodiment of the present invention, is attached to at least one otherend of said at least on wire. When activated the lightweightminiaturized circuitry 181003 configures waveforms that are directed tothe flexible coils (181001) to create PEMF signals that are coupled to atarget pathway structure.

Having described embodiments for an electromagnetic treatment inductiveapparatus and a method for using same, it is noted that modificationsand variations can be made by persons skilled in the art in light of theabove teachings. It is therefore to be understood that changes may bemade in the particular embodiments of the invention disclosed which arewithin the scope and spirit of the invention as defined by the appendedclaims.

Part 3

Induced time-varying currents from PEMF or PRF devices flow in a targetpathway structure such as a molecule, cell, tissue, and organ, and it isthese currents that are a stimulus to which cells and tissues can reactin a physiologically meaningful manner. The electrical properties of atarget pathway structure affect levels and distributions of inducedcurrent. Molecules, cells, tissue, and organs are all in an inducedcurrent pathway such as cells in a gap junction contact. Ion or ligandinteractions at binding sites on macromolecules that may reside on amembrane surface are voltage dependent processes, that iselectrochemical, that can respond to an induced electromagnetic field(“E”). Induced current arrives at these sites via a surrounding ionicmedium. The presence of cells in a current pathway causes an inducedcurrent (“J”) to decay more rapidly with time (“J(t)”). This is due toan added electrical impedance of cells from membrane capacitance andtime constants of binding and other voltage sensitive membrane processessuch as membrane transport.

Equivalent electrical circuit models representing various membrane andcharged interface configurations have been derived. For example, inCalcium (“Ca²⁺”) binding, the change in concentration of bound Ca²⁺ at abinding site due to induced E may be described in a frequency domain byan impedance expression such as:

${Z_{b}(\omega)} = {R_{ion} + \frac{1}{i\; \omega \; C_{ion}}}$

which has the form of a series resistance-capacitance electricalequivalent circuit. Where ω is angular frequency defined as 2πf, where fis frequency, i=−1^(1/2), Z_(b)(ω) is the binding impedance, and R_(ion)and C_(ion) are equivalent binding resistance and capacitance of an ionbinding pathway. The value of the equivalent binding time constant,τ_(ion)=R_(ion)C_(ion), is related to a ion binding rate constant,k_(b), via τ_(ion)=R_(ion)C_(ion)=1/k_(b). Thus, the characteristic timeconstant of this pathway is determined by ion binding kinetics.

Induced E from a PEMF or PRF signal can cause current to flow into anion binding pathway and affect the number of Ca²⁺ ions bound per unittime. An electrical equivalent of this is a change in voltage across theequivalent binding capacitance C_(ion), which is a direct measure of thechange in electrical charge stored by C_(ion). Electrical charge isdirectly proportional to a surface concentration of Ca²⁺ ions in thebinding site, that is storage of charge is equivalent to storage of ionsor other charged species on cell surfaces and junctions. Electricalimpedance measurements, as well as direct kinetic analyses of bindingrate constants, provide values for time constants necessary forconfiguration of a PMF waveform to match a bandpass of target pathwaystructures. This allows for a required range of frequencies for anygiven induced E waveform for optimal coupling to target impedance, suchas bandpass.

Ion binding to regulatory molecules is a frequent EMF target, forexample Ca²⁺ binding to calmodulin (“CaM”). Use of this pathway is basedupon acceleration of wound repair, for example bone repair, thatinvolves modulation of growth factors released in various stages ofrepair. Growth factors such as platelet derived growth factor (“PDGF”),fibroblast growth factor (“FGF”), and epidermal growth factor (“EGF”)are all involved at an appropriate stage of healing. Angiogenesis andneovascularization are also integral to wound repair and can bemodulated by PMF. All of these factors are Ca/CaM-dependent.

Utilizing a Ca/CaM pathway a waveform can be configured for whichinduced power is sufficiently above background thermal noise power.Under correct physiological conditions, this waveform can have aphysiologically significant bioeffect.

Application of a Power SNR model to Ca/CaM requires knowledge ofelectrical equivalents of Ca²⁺ binding kinetics at CaM. Within firstorder binding kinetics, changes in concentration of bound Ca²⁺ at CaMbinding sites over time may be characterized in a frequency domain by anequivalent binding time constant, τ_(ion)=R_(ion)C_(ion), where R_(ion)and C_(ion) are equivalent binding resistance and capacitance of the ionbinding pathway. τ_(ion) is related to a ion binding rate constant,k_(b), via τ_(ion)=R_(ion)C_(ion)=1/k_(b). Published values for k_(b)can then be employed in a cell array model to evaluate SNR by comparingvoltage induced by a PRF signal to thermal fluctuations in voltage at aCaM binding site. Employing numerical values for PMF response, such asV_(max)=6.5×10⁻⁷ sec⁻¹, =2.50/1, K_(D)=30 μM, [Ca²⁺CaM]=K_(D)(+[CaM]),yields k_(b)=665 sec⁻¹ (τ_(ion)=1.5 msec). Such a value for τ_(ion) canbe employed in an electrical equivalent circuit for ion binding whilepower SNR analysis can be performed for any waveform structure.

According to an embodiment of the present invention a mathematical modelcan be configured to assimilate that thermal noise is present in allvoltage dependent processes and represents a minimum thresholdrequirement to establish adequate SNR. Power spectral density, S_(n)(ω),of thermal noise can be expressed as:

S _(n)(ω)=4kT Re[Z _(M)(x,ω)]

where Z_(M)(x,ω) is electrical impedance of a target pathway structure,x is a dimension of a target pathway structure and Re denotes a realpart of impedance of a target pathway structure. Z_(M)(x,ω) can beexpressed as:

${Z_{M}( {x,\omega} )} = {\lbrack \frac{R_{e} + R_{i} + R_{g}}{\gamma} \rbrack {\tanh ( {\gamma \; x} )}}$

This equation clearly shows that electrical impedance of the targetpathway structure, and contributions from extracellular fluid resistance(“R_(e)”), intracellular fluid resistance (“R_(i)”) and intermembraneresistance (“R_(g)”) which are electrically connected to a targetpathway structure, all contribute to noise filtering.

A typical approach to evaluation of SNR uses a single value of a rootmean square (RMS) noise voltage. This is calculated by taking a squareroot of an integration of S_(n)(ω)=4 kT Re[z_(M) (x, ω)] over allfrequencies relevant to either complete membrane response, or tobandwidth of a target pathway structure. SNR can be expressed by aratio:

${SNR} = \frac{{V_{M}(\omega)}}{RMS}$

where |V_(M)(ω)| is maximum amplitude of voltage at each frequency asdelivered by a chosen waveform to the target pathway structure.

An embodiment according to the present invention comprises a pulse burstenvelope having a high spectral density, so that the effect of therapyupon the relevant dielectric pathways, such as, cellular membranereceptors, ion binding to cellular enzymes and general transmembranepotential changes, is enhanced. Accordingly by increasing a number offrequency components transmitted to relevant cellular pathways, a largerange of biophysical phenomena, such as modulating growth factor andcytokine release and ion binding at regulatory molecules, applicable toknown healing mechanisms is accessible. According to an embodiment ofthe present invention applying a random, or other high spectral densityenvelope, to a pulse burst envelope of mono- or bi-polar rectangular orsinusoidal pulses inducing peak electric fields between about 10⁻⁶ andabout 100 V/cm, produces a greater effect on biological healingprocesses applicable to both soft and hard tissues.

According to yet another embodiment of the present invention by applyinga high spectral density voltage envelope as a modulating or pulse-burstdefining parameter, power requirements for such amplitude modulatedpulse bursts can be significantly lower than that of an unmodulatedpulse burst containing pulses within a similar frequency range. This isdue to a substantial reduction in duty cycle within repetitive bursttrains brought about by imposition of an irregular, and preferablyrandom, amplitude onto what would otherwise be a substantially uniformpulse burst envelope. Accordingly, the dual advantages, of enhancedtransmitted dosimetry to the relevant dielectric pathways and ofdecreased power requirement are achieved.

Referring to FIG. 19, wherein FIG. 19 is a flow diagram of a method fordelivering electromagnetic signals to angiogenesis andneovascularization target pathway structures such as ions and ligands ofplants, animals, and humans for therapeutic and prophylactic purposesaccording to an embodiment of the present invention. A mathematicalmodel having at least one waveform parameter is applied to configure atleast one waveform to be coupled to a angiogenesis andneovascularization target pathway structure such as ions and ligands(Step 19101). The configured waveform satisfies a SNR or Power SNR modelso that for a given and known angiogenesis and neovascularization targetpathway structure it is possible to choose at least one waveformparameter so that a waveform is detectable in the angiogenesis andneovascularization target pathway structure above its backgroundactivity (Step 19102) such as baseline thermal fluctuations in voltageand electrical impedance at a target pathway structure that depend upona state of a cell and tissue, that is whether the state is at least oneof resting, growing, replacing, and responding to injury. A preferredembodiment of a generated electromagnetic signal is comprised of a burstof arbitrary waveforms having at least one waveform parameter thatincludes a plurality of frequency components ranging from about 0.01 Hzto about 100 MHz wherein the plurality of frequency components satisfiesa Power SNR model (Step 19102). A repetitive electromagnetic signal canbe generated for example inductively or capacitively, from saidconfigured at least one waveform (Step 19103). The electromagneticsignal is coupled to a angiogenesis and neovascularization targetpathway structure such as ions and ligands by output of a couplingdevice such as an electrode or an inductor, placed in close proximity tothe target pathway structure (Step 19104). The coupling enhancesmodulation of binding of ions and ligands to regulatory molecule inliving tissues and cells.

FIG. 20 illustrates a preferred embodiment of an apparatus according tothe present invention. A miniature control circuit 20201 is coupled toan end of at least one connector 20202 such as wire. The opposite end ofthe at least one connector is coupled to a generating device such as apair of electrical coils 20203. The miniature control circuit 20201 isconstructed in a manner that applies a mathematical model that is usedto configure waveforms. The configured waveforms have to satisfy a SNRor Power SNR model so that for a given and known angiogenesis andneovascularization target pathway structure, it is possible to choosewaveform parameters that satisfy SNR or Power SNR so that a waveform isdetectable in the angiogenesis and neovascularization target pathwaystructure above its background activity. A preferred embodimentaccording to the present invention applies a mathematical model toinduce a time-varying magnetic field and a time-varying electric fieldin a angiogenesis and neovascularization target pathway structure suchas ions and ligands, comprising about 10 to about 100 msec bursts ofabout 1 to about 100 microsecond rectangular pulses repeating at about0.1 to about 10 pulses per second. Peak amplitude of the inducedelectric field is between about 1 uV/cm and about 100 mV/cm, variedaccording to a modified 1/f function where f=frequency. A waveformconfigured using a preferred embodiment according to the presentinvention may be applied to a angiogenesis and neovascularization targetpathway structure such as ions and ligands for a preferred totalexposure time of under 1 minute to 240 minutes daily. However otherexposure times can be used. Waveforms configured by the miniaturecontrol circuit 20201 are directed to a generating device 20203 such aselectrical coils via connector 20202. The generating device 20203delivers a pulsing magnetic field configured according to a mathematicalmodel, that can be used to provide treatment to a angiogenesis andneovascularization target pathway structure such as a heart in a chest20204. The miniature control circuit applies a pulsing magnetic fieldfor a prescribed time and can automatically repeat applying the pulsingmagnetic field for as many applications as are needed in a given timeperiod, for example 10 times a day. A preferred embodiment according tothe present invention can be positioned to treat the heart in a chest20204 by a positioning device. Coupling a pulsing magnetic field to aangiogenesis and neovascularization target pathway structure such asions and ligands, therapeutically and prophylactically reducesinflammation thereby reducing pain and promotes healing. When electricalcoils are used as the generating device 20203, the electrical coils canbe powered with a time varying magnetic field that induces a timevarying electric field in a target pathway structure according toFaraday's law. An electromagnetic signal generated by the generatingdevice 20203 can also be applied using electrochemical coupling, whereinelectrodes are in direct contact with skin or another outer electricallyconductive boundary of a target pathway structure. Yet in anotherembodiment according to the present invention, the electromagneticsignal generated by the generating device 20203 can also be appliedusing electrostatic coupling wherein an air gap exists between agenerating device 20203 such as an electrode and a angiogenesis andneovascularization target pathway structure such as ions and ligands. Anadvantage of the preferred embodiment according to the present inventionis that its ultra lightweight coils and miniaturized circuitry allow foruse with common physical therapy treatment modalities and at any bodylocation for which pain relief and healing is desired. An advantageousresult of application of the preferred embodiment according to thepresent invention is that a living organism's angiogenesis andneovascularization can be maintained and enhanced.

FIG. 21 depicts a block diagram of a preferred embodiment according tothe present invention of a miniature control circuit 21300. Theminiature control circuit 21300 produces waveforms that drive agenerating device such as wire coils described above in FIG. 20. Theminiature control circuit can be activated by any activation means suchas an on/off switch. The miniature control circuit 21300 has a powersource such as a lithium battery 21301. A preferred embodiment of thepower source has an output voltage of 3.3 V but other voltages can beused. In another embodiment according to the present invention the powersource can be an external power source such as an electric currentoutlet such as an AC/DC outlet, coupled to the present invention forexample by a plug and wire. A switching power supply 21302 controlsvoltage to a micro-controller 21303. A preferred embodiment of themicro-controller 21303 uses an 8 bit 4 MHz micro-controller 21303 butother bit MHz combination micro-controllers may be used. The switchingpower supply 21302 also delivers current to storage capacitors 21304. Apreferred embodiment of the present invention uses storage capacitorshaving a 220 uF output but other outputs can be used. The storagecapacitors 21304 allow high frequency pulses to be delivered to acoupling device such as inductors (Not Shown). The micro-controller21303 also controls a pulse shaper 21305 and a pulse phase timingcontrol 21306. The pulse shaper 21305 and pulse phase timing control 306determine pulse shape, burst width, burst envelope shape, and burstrepetition rate. An integral waveform generator, such as a sine wave orarbitrary number generator can also be incorporated to provide specificwaveforms. A voltage level conversion sub-circuit 21308 controls aninduced field delivered to a target pathway structure. A switchingHexfet 21308 allows pulses of randomized amplitude to be delivered tooutput 21309 that routes a waveform to at least one coupling device suchas an inductor. The micro-controller 21303 can also control totalexposure time of a single treatment of a target pathway structure suchas a molecule, cell, tissue, and organ. The miniature control circuit21300 can be constructed to apply a pulsing magnetic field for aprescribed time and to automatically repeat applying the pulsingmagnetic field for as many applications as are needed in a given timeperiod, for example 10 times a day. A preferred embodiment according tothe present invention uses treatments times of about 10 minutes to about30 minutes.

Referring to FIG. 22 an embodiment according to the present invention ofa waveform 22400 is illustrated. A pulse 22401 is repeated within aburst 22402 that has a finite duration 22403. The duration 22403 is suchthat a duty cycle which can be defined as a ratio of burst duration tosignal period is between about 1 to about 10⁻⁵. A preferred embodimentaccording to the present invention utilizes pseudo rectangular 10microsecond pulses for pulse 22401 applied in a burst 22402 for about 10to about 50 msec having a modified 1/f amplitude envelope 22404 and witha finite duration 22403 corresponding to a burst period of between about0.1 and about 10 seconds.

Example 1

The Power SNR approach for PMF signal configuration has been testedexperimentally on calcium dependent myosin phosphorylation in a standardenzyme assay. The cell-free reaction mixture was chosen forphosphorylation rate to be linear in time for several minutes, and forsub-saturation Ca²⁺ concentration. This opens the biological window forCa²⁺/CaM to be EMF-sensitive. This system is not responsive to PMF atlevels utilized in this study if Ca²⁺ is at saturation levels withrespect to CaM, and reaction is not slowed to a minute time range.Experiments were performed using myosin light chain (“MLC”) and myosinlight chain kinase (“MLCK”) isolated from turkey gizzard. A reactionmixture consisted of a basic solution containing 40 mM Hepes buffer, pH7.0; 0.5 mM magnesium acetate; 1 mg/ml bovine serum albumin, 0.1% (w/v)Tween 80; and 1 mM EGTA12. Free Ca²⁺ was varied in the 1-7 μM range.Once Ca²⁺ buffering was established, freshly prepared 70 nM CaM, 160 nMMLC and 2 nM MLCK were added to the basic solution to form a finalreaction mixture. The low MLC/MLCK ratio allowed linear time behavior inthe minute time range. This provided reproducible enzyme activities andminimized pipetting time errors.

The reaction mixture was freshly prepared daily for each series ofexperiments and was aliquoted in 100 μL portions into 1.5 ml Eppendorftubes. All Eppendorf tubes containing reaction mixture were kept at 0°C. then transferred to a specially designed water bath maintained at37±0.1° C. by constant perfusion of water prewarmed by passage through aFisher Scientific model 900 heat exchanger. Temperature was monitoredwith a thermistor probe such as a Cole-Parmer model 8110-20, immersed inone Eppendorf tube during all experiments. Reaction was initiated with2.5 μM 32P ATP, and was stopped with Laemmli Sample Buffer solutioncontaining 30 μM EDTA. A minimum of five blank samples were counted ineach experiment. Blanks comprised a total assay mixture minus one of theactive components Ca²⁺, CaM, MLC or MLCK. Experiments for which blankcounts were higher than 300 cpm were rejected. Phosphorylation wasallowed to proceed for 5 min and was evaluated by counting 32Pincorporated in MLC using a TM Analytic model 5303 Mark V liquidscintillation counter.

The signal comprised repetitive bursts of a high frequency waveform.Amplitude was maintained constant at 0.2 G and repetition rate was 1burst/sec for all exposures. Burst duration varied from 65 μsec to 1000μsec based upon projections of Power SNR analysis which showed thatoptimal Power SNR would be achieved as burst duration approached 500μsec. The results are shown in FIG. 7 wherein burst width 701 in μsec isplotted on the x-axis and Myosin Phosphorylation 702 as treated/sham isplotted on the y-axis. It can be seen that the PMF effect on Ca²⁺binding to CaM approaches its maximum at approximately 500 μsec, just asillustrated by the Power SNR model.

These results confirm that a PMF signal, configured according to anembodiment of the present invention, would maximally increase myosinphosphorylation for burst durations sufficient to achieve optimal PowerSNR for a given magnetic field amplitude.

Example 2

According to an embodiment of the present invention use of a Power SNRmodel was further verified in an in vivo wound repair model. A rat woundmodel has been well characterized both biomechanically andbiochemically, and was used in this study. Healthy, young adult maleSprague Dawley rats weighing more than 300 grams were utilized.

The animals were anesthetized with an intraperitoneal dose of Ketamine75 mg/kg and Medetomidine 0.5 mg/kg. After adequate anesthesia had beenachieved, the dorsum was shaved, prepped with a dilute betadine/alcoholsolution, and draped using sterile technique. Using a #10 scalpel, an8-cm linear incision was performed through the skin down to the fasciaon the dorsum of each rat. The wound edges were bluntly dissected tobreak any remaining dermal fibers, leaving an open wound approximately 4cm in diameter. Hemostasis was obtained with applied pressure to avoidany damage to the skin edges. The skin edges were then closed with a 4-0Ethilon running suture. Post-operatively, the animals receivedBuprenorphine 0.1-0.5 mg/kg, intraperitoneal. They were placed inindividual cages and received food and water ad libitum.

PMF exposure comprised two pulsed radio frequency waveforms. The firstwas a standard clinical PRF signal comprising a 65 μsec burst of 27.12MHz sinusoidal waves at 1 Gauss amplitude and repeating at 600bursts/sec. The second was a PRF signal reconfigured according to anembodiment of the present invention. For this signal burst duration wasincreased to 2000 μsec and the amplitude and repetition rate werereduced to 0.2 G and 5 bursts/sec respectively. PRF was applied for 30minutes twice daily.

Tensile strength was performed immediately after wound excision. Two 1cm width strips of skin were transected perpendicular to the scar fromeach sample and used to measure the tensile strength in kg/mm². Thestrips were excised from the same area in each rat to assure consistencyof measurement. The strips were then mounted on a tensiometer. Thestrips were loaded at 10 mm/min and the maximum force generated beforethe wound pulled apart was recorded. The final tensile strength forcomparison was determined by taking the average of the maximum load inkilograms per mm² of the two strips from the same wound.

The results showed average tensile strength for the 65 μsec 1 Gauss PRFsignal was 19.3±4.3 kg/mm² for the exposed group versus 13.0±3.5 kg/mm²for the control group (p<0.01), which is a 48% increase. In contrast,the average tensile strength for the 2000 μsec 0.2 Gauss PRF signal,configured according to an embodiment of the present invention using aPower SNR model was 21.2±5.6 kg/mm² for the treated group versus13.7±4.1 kg/mm² (p<0.01) for the control group, which is a 54% increase.The results for the two signals were not significantly different fromeach other.

These results demonstrate that an embodiment of the present inventionallowed a new PRF signal to be configured that could be produced withsignificantly lower power. The PRF signal configured according to anembodiment of the present invention, accelerated wound repair in the ratmodel in a low power manner versus that for a clinical PRF signal whichaccelerated wound repair but required more than two orders of magnitudemore power to produce.

Example 3

In this example Jurkat cells react to PMF stimulation of a T-cellreceptor with cell cycle arrest and thus behave like normalT-lymphocytes stimulated by antigens at the T-cell receptor such asanti-CD3. For example in bone healing, results have shown both 60 Hz andPEMF fields decrease DNA synthesis of Jurkat cells, as is expected sincePMF interacts with the T-cell receptor in the absence of a costimulatorysignal. This is consistent with an anti-inflammatory response, as hasbeen observed in clinical applications of PMF stimuli. The PEMF signalis more effective. A dosimetry analysis performed according to anembodiment of the present invention demonstrates why both signals areeffective and why PEMF signals have a greater effect than 60 Hz signalson Jurkat cells in the most EMF-sensitive growth stage.

Comparison of dosimetry from the two signals employed involvesevaluation of the ratio of the Power spectrum of the thermal noisevoltage that is Power SNR, to that of the induced voltage at theEMF-sensitive target pathway structure. The target pathway structureused is ion binding at receptor sites on Jurkat cells suspended in 2 mmof culture medium. The average peak electric field at the binding sitefrom a PEMF signal comprising 5 msec burst of 200 μsec pulses repeatingat 15/sec, was 1 mV/cm, while for a 60 Hz signal it was 50 μV/cm.

Example 4

In this example electromagnetic field energy was used to stimulateneovascularization in an in vivo model. Two different signal wereemployed, one configured according to prior art and a second configuredaccording to an embodiment of the present invention.

One hundred and eight Sprague-Dawley male rats weighing approximately300 grams each, were equally divided into nine groups. All animals wereanesthetized with a mixture of ketamine/acepromazine/Stadol at 0.1 cc/g.Using sterile surgical techniques, each animal had a 12 cm to 14 cmsegment of tail artery harvested using microsurgical technique. Theartery was flushed with 60 U/ml of heparinized saline to remove anyblood or emboli.

These tail vessels, with an average diameter of 0.4 mm to 0.5 mm, werethen sutured to the transected proximal and distal segments of the rightfemoral artery using two end-to-end anastomoses, creating a femoralarterial loop. The resulting loop was then placed in a subcutaneouspocket created over the animal's abdominal wall/groin musculature, andthe groin incision was closed with 4-0 Ethilon. Each animal was thenrandomly placed into one of nine groups: groups 1 to 3 (controls), theserats received no electromagnetic field treatments and were killed at 4,8, and 12 weeks; groups 4 to 6, 30 min. treatments twice a day using 0.1gauss electromagnetic fields for 4, 8, and 12 weeks (animals were killedat 4, 8, and 12 weeks, respectively); and groups 7 to 9, 30 min.treatments twice a day using 2.0 gauss electromagnetic fields for 4, 8,and 12 weeks (animals were killed at 4, 8, and 12 weeks, respectively).

Pulsed electromagnetic energy was applied to the treated groups using adevice constructed according to an embodiment of the present invention.Animals in the experimental groups were treated for 30 minutes twice aday at either 0.1 gauss or 2.0 gauss, using short pulses (2 msec to 20msec) 27.12 MHz. Animals were positioned on top of the applicator headand confined to ensure that treatment was properly applied. The ratswere reanesthetized with ketamine/acepromazine/Stadol intraperitoneallyand 100 U/kg of heparin intravenously. Using the previous groinincision, the femoral artery was identified and checked for patency. Thefemoral/tail artery loop was then isolated proximally and distally fromthe anastomoses sites, and the vessel was clamped off. Animals were thenkilled. The loop was injected with saline followed by 0.5 cc to 1.0 ccof colored latex through a 25-gauge cannula and clamped. The overlyingabdominal skin was carefully resected, and the arterial loop wasexposed. Neovascularization was quantified by measuring the surface areacovered by new blood-vessel formation delineated by the intraluminallatex. All results were analyzed using the SPSS statistical analysispackage.

The most noticeable difference in neovascularization between treatedversus untreated rats occurred at week 4. At that time, no new vesselformation was found among controls, however, each of the treated groupshad similar statistically significant evidence of neovascularization at0 cm2 versus 1.42±0.80 cm2 (p<0.001). These areas appeared as a latexblush segmentally distributed along the sides of the arterial loop. At 8weeks, controls began to demonstrate neovascularization measured at0.7±0.82 cm2. Both treated groups at 8 weeks again had approximatelyequal statistically significant (p<0.001) outcroppings of blood vesselsof 3.57±1.82 cm2 for the 0.1 gauss group and of 3.77±1.82 cm2 for the2.0 gauss group. At 12 weeks, animals in the control group displayed1.75±0.95 cm2 of neovascularization, whereas the 0.1 gauss groupdemonstrated 5.95±3.25 cm2, and the 2.0 gauss group showed 6.20±3.95 cm2of arborizing vessels. Again, both treated groups displayed comparablestatistically significant findings (p<0.001) over controls.

These experimental findings demonstrate that electromagnetic fieldstimulation of an isolated arterial loop according to an embodiment ofthe present invention increases the amount of quantifiableneovascularization in an in vivo rat model. Increased angiogenesis wasdemonstrated in each of the treated groups at each of the sacrificedates. No differences were found between the results of the two gausslevels tested as predicted by the teachings of the present invention.

Having described embodiments for an apparatus and a method fordelivering electromagnetic treatment to human, animal and plantmolecules, cells, tissue and organs, it is noted that modifications andvariations can be made by persons skilled in the art in light of theabove teachings. It is therefore to be understood that changes may bemade in the particular embodiments of the invention disclosed which arewithin the scope and spirit of the invention as defined by the appendedclaims.

Part 4

An embodiment according to the present invention provides a higherspectral density to a pulse burst envelope resulting in enhancedeffectiveness of therapy upon relevant dielectric pathways, such as,cellular membrane receptors, ion binding to cellular enzymes and generaltransmembrane potential changes. An embodiment according to the presentinvention increases the number of frequency components transmitted torelevant cellular pathways, thereby providing access to a larger rangeof biophysical phenomena applicable to known healing mechanisms, forexample modulation of growth factor and cytokine release, and ionbinding at regulatory molecules. By applying a random, or other highspectral density envelope, according to a mathematical model defined bySNR or Power SNR in a transduction pathway, to a pulse burst envelope ofmono- or bi-polar rectangular or sinusoidal pulses inducing peakelectric fields between 10⁻⁶ and 10 volts per centimeter (V/cm), agreater effect could be accomplished on biological healing processesapplicable to both soft and hard tissues.

An advantageous result of the present invention, is that by applying ahigh spectral density voltage envelope as the modulating or pulse-burstdefining parameter, according to a mathematical model defined by SNR orPower SNR in a transduction pathway, the power requirement for suchamplitude modulated pulse bursts can be significantly lower than that ofan unmodulated pulse burst containing pulses within the same frequencyrange. Accordingly, the advantages of enhanced transmitted dosimetry tothe relevant dielectric target pathways and of decreased powerrequirement are achieved. Another advantage of the present invention isthe acceleration of wound repair.

Known mechanisms of wound repair involve the naturally timed release ofthe appropriate growth factor or cytokine in each stage of wound repairas applied to humans, animals and plants. Specifically, wound repairinvolves an inflammatory phase, angiogenesis, cell proliferation,collagen production, and remodeling stages. There are timed releases ofspecific cytokines and growth factors in each stage. Electromagneticfields are known to enhance blood flow and to enhance the binding ofions which, in turn, can accelerate each healing phase. It is an objectof this invention to provide an improved means to enhance the action andaccelerate the intended effects or improve efficacy as well as othereffects of the cytokines and growth factors relevant to each stage ofwound repair.

Induced time-varying currents from PEMF or PRF devices flow in a targetpathway structure such as a molecule, cell, tissue, and organ, and it isthese currents that are a stimulus to which cells and tissues can reactin a physiologically meaningful manner. The electrical properties of atarget pathway structure affect levels and distributions of inducedcurrent. Molecules, cells, tissue, and organs are all in an inducedcurrent pathway such as cells in a gap junction contact. Ion or ligandinteractions at binding sites on macromolecules that may reside on amembrane surface are voltage dependent processes, for exampleelectrochemical, that can respond to an induced electromagnetic field(“E”). Induced current arrives at these sites via a surrounding ionicmedium. The presence of cells in a current pathway causes an inducedcurrent (“J”) to decay more rapidly with time (“J(t)”). This is due toan added electrical impedance of cells from membrane capacitance andtime constants of binding and other voltage sensitive membrane processessuch as membrane transport.

Equivalent electrical circuit models representing various membrane andcharged interface configurations have been derived. For example, inCalcium (“Ca²⁺”) binding, the change in concentration of bound Ca²⁺ at abinding site due to induced E may be described in a frequency domain byan impedance expression such as:

${Z_{b}(\omega)} = {R_{ion} + \frac{1}{i\; \omega \; C_{ion}}}$

which has the form of a series resistance-capacitance electricalequivalent circuit. Where ω is angular frequency defined as 2πf, where fis frequency, i=−1^(1/2), Z_(b)(ω) is the binding impedance, and R_(ion)and C_(ion) are equivalent binding resistance and capacitance of an ionbinding pathway. The value of the equivalent binding time constant,τ_(ion)=R_(ion)C_(ion), is related to a ion binding rate constant,k_(b), via τ_(ion)=R_(ion)C_(ion)=1/k_(b). Thus, the characteristic timeconstant of this pathway is determined by ion binding kinetics.

Induced E from a PEMF or PRF signal can cause current to flow into anion binding pathway and affect the number of Ca²⁺ ions bound per unittime. An electrical equivalent of this is a change in voltage across theequivalent binding capacitance C_(ion), which is a direct measure of thechange in electrical charge stored by C_(ion). Electrical charge isdirectly proportional to a surface concentration of Ca²⁺ ions in thebinding site, that is storage of charge is equivalent to storage of ionsor other charged species on cell surfaces and junctions. Electricalimpedance measurements, as well as direct kinetic analyses of bindingrate constants, provide values for time constants necessary forconfiguration of a PMF waveform to match a bandpass of target pathwaystructures. This allows for a required range of frequencies for anygiven induced E waveform for optimal coupling to target impedance, suchas bandpass.

Ion binding to regulatory molecules is a frequent EMF target, forexample Ca²⁺ binding to calmodulin (“CaM”). Use of this pathway is basedupon acceleration of tissue repair, for example bone repair, woundrepair, hair repair, and repair of molecules, cells, tissues, and organsthat involves modulation of growth factors released in various stages ofrepair. Growth factors such as platelet derived growth factor (“PDGF”),fibroblast growth factor (“FGF”), and epidermal growth factor (“EGF”)are all involved at an appropriate stage of healing. Angiogenesis andneovascularization are also integral to tissue growth and repair and canbe modulated by PMF. All of these factors are Ca/CaM-dependent.

Utilizing a Ca/CaM pathway a waveform can be configured for whichinduced power is sufficiently above background thermal noise power.Under correct physiological conditions, this waveform can have aphysiologically significant bioeffect.

Application of a Power SNR model to Ca/CaM requires knowledge ofelectrical equivalents of Ca²⁺ binding kinetics at CaM. Within firstorder binding kinetics, changes in concentration of bound Ca²⁺ at CaMbinding sites over time may be characterized in a frequency domain by anequivalent binding time constant, τ_(ion)=R_(ion)C_(ion), where R_(ion)and C_(ion) are equivalent binding resistance and capacitance of the ionbinding pathway. τ_(ion) is related to a ion binding rate constant,k_(b), via τ_(ion)=R_(ion)C_(ion)=1/k_(b). Published values for k_(b)can then be employed in a cell array model to evaluate SNR by comparingvoltage induced by a PRF signal to thermal fluctuations in voltage at aCaM binding site. Employing numerical values for PMF response, such asV_(max)=6.5×10⁻⁷ sec⁻¹, =2.50/1, K_(D)=30 μM, [Ca²⁺CaM]=K_(D)(+[CaM]),yields k_(b)=665 sec⁻¹ (τ_(ion)=1.5 msec). Such a value for τ_(ion) canbe employed in an electrical equivalent circuit for ion binding whilepower SNR analysis can be performed for any waveform structure.

According to an embodiment of the present invention a mathematical modelfor example a mathematical equation and or a series of mathematicalequations can be configured to assimilate that thermal noise is presentin all voltage dependent processes and represents a minimum thresholdrequirement to establish adequate SNR. For example a mathematical modelthat represents a minimum threshold requirement to establish adequateSNR can be configured to include power spectral density of thermal noisesuch that power spectral density, S_(n)(ω), of thermal noise can beexpressed as:

S _(n)(ω)=4kT Re[Z _(M)(x,ω)]

where Z_(M)(x,ω) is electrical impedance of a target pathway structure,x is a dimension of a target pathway structure and Re denotes a realpart of impedance of a target pathway structure. Z_(M)(x,ω) can beexpressed as:

${Z_{M}( {x,\omega} )} = {\lbrack \frac{R_{e} + R_{i} + R_{g}}{\gamma} \rbrack {\tanh ( {\gamma \; x} )}}$

This equation clearly shows that electrical impedance of the targetpathway structure, and contributions from extracellular fluid resistance(“R_(e)”), intracellular fluid resistance (“R_(i)”) and intermembraneresistance (“R_(g)”) which are electrically connected to a targetpathway structures, all contribute to noise filtering.

A typical approach to evaluation of SNR uses a single value of a rootmean square (RMS) noise voltage. This is calculated by taking a squareroot of an integration of

S_(n) (ω)=4 kT Re[Z_(M) (x, ω)] over all frequencies relevant to eithercomplete membrane response, or to bandwidth of a target pathwaystructure. SNR can be expressed by a ratio:

${SNR} = \frac{{V_{M}(\omega)}}{RMS}$

where |V_(M)(ω)| is maximum amplitude of voltage at each frequency asdelivered by a chosen waveform to the target pathway structure.

An embodiment according to the present invention comprises a pulse burstenvelope having a high spectral density, so that the effect of therapyupon the relevant dielectric pathways, such as, cellular membranereceptors, ion binding to cellular enzymes and general transmembranepotential changes, is enhanced. Accordingly by increasing a number offrequency components transmitted to relevant cellular pathways, a largerange of biophysical phenomena, such as modulating growth factor andcytokine release and ion binding at regulatory molecules, applicable toknown tissue growth mechanisms is accessible. According to an embodimentof the present invention applying a random, or other high spectraldensity envelope, to a pulse burst envelope of mono- or bi-polarrectangular or sinusoidal pulses inducing peak electric fields betweenabout 10⁻⁸ and about 100 V/cm, produces a greater effect on biologicalhealing processes applicable to both soft and hard tissues.

According to yet another embodiment of the present invention by applyinga high spectral density voltage envelope as a modulating or pulse-burstdefining parameter, power requirements for such amplitude modulatedpulse bursts can be significantly lower than that of an unmodulatedpulse burst containing pulses within a similar frequency range. This isdue to a substantial reduction in duty cycle within repetitive bursttrains brought about by imposition of an irregular, and preferablyrandom, amplitude onto what would otherwise be a substantially uniformpulse burst envelope. Accordingly, the dual advantages, of enhancedtransmitted dosimetry to the relevant dielectric pathways and ofdecreased power requirement are achieved.

Referring to FIG. 23, wherein FIG. 23 is a flow diagram of a methodaccording to an embodiment of the present invention, for acceleratingwound repair by delivering electromagnetic signals that can be pulsed,to target pathway structures such as ions and ligands of animals andhumans, for therapeutic and prophylactic purposes. Target pathwaystructures can also include but are not limited to tissues, cells,organs, and molecules.

Configuring at least one waveform having at least one waveform parameterto be coupled to the target pathway structure such as ions and ligands(Step 23101).

The at least one waveform parameter is selected to maximize at least oneof a signal to noise ratio and a Power Signal to Noise ratio in a targetpathway structure so that a waveform is detectable in the target pathwaystructure above its background activity (Step 23102) such as baselinethermal fluctuations in voltage and electrical impedance at a targetpathway structure that depend upon a state of a cell and tissue, that iswhether the state is at least one of resting, growing, replacing, andresponding to injury to produce physiologically beneficial results. Tobe detectable in the target pathway structure the value of said at leastone waveform parameter is chosen by using a constant of said targetpathway structure to evaluate at least one of a signal to noise ratio,and a Power signal to noise ratio, to compare voltage induced by said atleast one waveform in said target pathway structure to baseline thermalfluctuations in voltage and electrical impedance in said target pathwaystructure whereby bioeffective modulation occurs in said target pathwaystructure by said at least one waveform by maximizing said at least oneof signal to noise ratio and Power signal to noise ratio, within abandpass of said target pathway structure.

A preferred embodiment of a generated electromagnetic signal iscomprised of a burst of arbitrary waveforms having at least one waveformparameter that includes a plurality of frequency components ranging fromabout 0.01 Hz to about 100 MHz wherein the plurality of frequencycomponents satisfies a Power SNR model (Step 23103). A repetitiveelectromagnetic signal can be generated for example inductively orcapacitively, from said configured at least one waveform (Step 23104).The electromagnetic signal can also be non-repetitive. Theelectromagnetic signal is coupled to a target pathway structure such asions and ligands by output of a coupling device such as an electrode oran inductor, placed in close proximity to the target pathway structure(Step 23105). The coupling enhances blood flow and modulation of bindingof ions and ligands to regulatory molecules in molecules, tissues,cells, and organs thereby accelerating wound repair.

FIG. 24 illustrates a preferred embodiment of an apparatus according tothe present invention. The apparatus is self-contained, lightweight, andportable. A miniature control circuit 24201 is coupled to an end of atleast one connector 24202 such as wire however the control circuit canalso operate wirelessly. The opposite end of the at least one connectoris coupled to a generating device such as an electrical coil 24203. Theminiature control circuit 24201 is constructed in a manner that appliesa mathematical model that is used to configure waveforms. The configuredwaveforms have to satisfy Power SNR so that for a given and known targetpathway structure, it is possible to choose waveform parameters thatsatisfy Power SNR so that a waveform produces physiologically beneficialresults, for example bioeffective modulation, and is detectable in thetarget pathway structure above its background activity. A preferredembodiment according to the present invention applies a mathematicalmodel to induce a time-varying magnetic field and a time-varyingelectric field in a target pathway structure such as ions and ligands,comprising about 0.1 to about 100 msec bursts of about 1 to about 100microsecond rectangular pulses repeating at about 0.1 to about 100pulses per second. Peak amplitude of the induced electric field isbetween about 1 uV/cm and about 100 mV/cm, varied according to amodified 1/f function where f=frequency. A waveform configured using apreferred embodiment according to the present invention may be appliedto a target pathway structure such as ions and ligands for a preferredtotal exposure time of under 1 minute to 240 minutes daily. Howeverother exposure times can be used. Waveforms configured by the miniaturecontrol circuit 24201 are directed to a generating device 24203 such aselectrical coils via connector 24202. The generating device 24203delivers a pulsing magnetic field that can be used to provide treatmentto a target pathway structure such as tissue. The miniature controlcircuit applies a pulsing magnetic field for a prescribed time and canautomatically repeat applying the pulsing magnetic field for as manyapplications as are needed in a given time period, for example 10 timesa day. The miniature control circuit can be configured to beprogrammable applying pulsing magnetic fields for any time repetitionsequence. A preferred embodiment according to the present invention canaccelerate wound repair by being incorporated into a positioning device24204, for example a bed. Coupling a pulsing magnetic field to a targetpathway structure such as ions and ligands, therapeutically andprophylactically reduces inflammation thereby advantageously reducingpain, promoting healing in targeted areas. When electrical coils areused as the generating device 24203, the electrical coils can be poweredwith a time varying magnetic field that induces a time varying electricfield in a target pathway structure according to Faraday's law. Anelectromagnetic signal generated by the generating device 24203 can alsobe applied using electrochemical coupling, wherein electrodes are indirect contact with skin or another outer electrically conductiveboundary of a target pathway structure. Yet in another embodimentaccording to the present invention, the electromagnetic signal generatedby the generating device 24203 can also be applied using electrostaticcoupling wherein an air gap exists between a generating device 24203such as an electrode and a target pathway structure such as ions andligands. An advantage of the preferred embodiment according to thepresent invention is that its ultra lightweight coils and miniaturizedcircuitry allow for use with common physical therapy treatmentmodalities and at any for which growth, pain relief, and tissue andorgan healing is desired. An advantageous result of application of thepreferred embodiment according to the present invention is that tissuegrowth, repair, and maintenance can be accomplished and enhancedanywhere and at anytime, for example while driving a car or watchingtelevision. Yet another advantageous result of application of thepreferred embodiment is that growth, repair, and maintenance ofmolecules, cells, tissues, and organs can be accomplished and enhancedanywhere and at anytime, for example while driving a car or watchingtelevision.

FIG. 25 depicts a block diagram of a preferred embodiment according tothe present invention of a miniature control circuit 25300. Theminiature control circuit 25300 produces waveforms that drive agenerating device such as wire coils described above in FIG. 24. Theminiature control circuit can be activated by any activation means suchas an on/off switch. The miniature control circuit 25300 has a powersource such as a lithium battery 25301. A preferred embodiment of thepower source has an output voltage of 3.3 V but other voltages can beused. In another embodiment according to the present invention the powersource can be an external power source such as an electric currentoutlet such as an AC/DC outlet, coupled to the present invention forexample by a plug and wire. A switching power supply 25302 controlsvoltage to a micro-controller 25303. A preferred embodiment of themicro-controller 25303 uses an 8 bit 4 MHz micro-controller 25303 butother bit MHz combination micro-controllers may be used. The switchingpower supply 25302 also delivers current to storage capacitors 25304. Apreferred embodiment of the present invention uses storage capacitorshaving a 220 uF output but other outputs can be used. The storagecapacitors 25304 allow high frequency pulses to be delivered to acoupling device such as inductors (Not Shown). The micro-controller25303 also controls a pulse shaper 25305 and a pulse phase timingcontrol 25306. The pulse shaper 25305 and pulse phase timing control25306 determine pulse shape, burst width, burst envelope shape, andburst repetition rate. An integral waveform generator, such as a sinewave or arbitrary number generator can also be incorporated to providespecific waveforms. A voltage level conversion sub-circuit 25307controls an induced field delivered to a target pathway structure. Aswitching Hexfet 25308 allows pulses of randomized amplitude to bedelivered to output 25309 that routes a waveform to at least onecoupling device such as an inductor. The micro-controller 25303 can alsocontrol total exposure time of a single treatment of a target pathwaystructure such as a molecule, cell, tissue, and organ. The miniaturecontrol circuit 25300 can be constructed to be programmable and apply apulsing magnetic field for a prescribed time and to automatically repeatapplying the pulsing magnetic field for as many applications as areneeded in a given time period, for example 10 times a day. A preferredembodiment according to the present invention uses treatments times ofabout 10 minutes to about 30 minutes.

Referring to FIGS. 26A and 26B a preferred embodiment according to thepresent invention of a coupling device 26400 such as an inductor isshown. The coupling device 26400 can be an electric coil 26401 woundwith single or multistrand flexible wire 26402 however solid wire canalso be used. In a preferred embodiment according to the presentinvention the wire is made of copper but other materials can be used.The multistrand flexible magnetic wire 26402 enables the electric coil26401 to conform to specific anatomical configurations such as a limb orjoint of a human or animal. A preferred embodiment of the electric coil26401 comprises about 1 to about 1000 turns of about 0.01 mm to about0.1 mm diameter at least one of single magnet wire and multistrandmagnet wire, wound on an initially circular form having an outerdiameter between about 2.5 cm and about 50 cm but other numbers of turnsand wire diameters can be used. A preferred embodiment of the electriccoil 26401 can be encased with a non-toxic PVC mould 403 but othernon-toxic moulds can also be used. The electric coil can also beincorporated in dressings, bandages, garments, and other structurestypically used for wound treatment.

Referring to FIG. 27 an embodiment according to the present invention ofa waveform 27500 is illustrated. A pulse 27501 is repeated within aburst 27502 that has a finite duration 27503. The duration 27503 is suchthat a duty cycle which can be defined as a ratio of burst duration tosignal period is between about 1 to about 10⁻⁵. A preferred embodimentaccording to the present invention utilizes pseudo rectangular 10microsecond pulses for pulse 27501 applied in a burst 27502 for about 10to about 50 msec having a modified 1/f amplitude envelope 27504 and witha finite duration 27503 corresponding to a burst period of between about0.1 and about 10 seconds but other waveforms, envelopes, and burstperiods may be used that conform to a mathematical model such as SNR andPower SNR.

FIG. 28 illustrates a preferred embodiment according to the presentinvention of a positioning device such as a wrist support. A positioningdevice 28600 such as a wrist support 28601 is worn on a human wrist28602. The positioning device can be constructed to be portable, can beconstructed to be disposable, and can be constructed to be implantable.The positioning device can be used in combination with the presentinvention in a plurality of ways, for example incorporating the presentinvention into the positioning device for example by stitching, affixingthe present invention onto the positioning device for example byVelcro®, and holding the present invention in place by constructing thepositioning device to be elastic.

In another embodiment according to the present invention, the presentinvention can be constructed as a stand-alone device of any size with orwithout a positioning device, to be used anywhere for example at home,at a clinic, at a treatment center, and outdoors. The wrist support28601 can be made with any anatomical and support material, such asneoprene. Coils 28603 are integrated into the wrist support 28601 suchthat a signal configured according to the present invention, for examplethe waveform depicted in FIG. 27, is applied from a dorsal portion thatis the top of the wrist to a plantar portion that is the bottom of thewrist. Micro-circuitry 28604 is attached to the exterior of the wristsupport 28601 using a fastening device such as Velcro® (Not Shown). Themicro-circuitry is coupled to one end of at least one connecting devicesuch as a flexible wire 28605. The other end of the at least oneconnecting device is coupled to the coils 28603. Other embodimentsaccording to the present invention of the positioning device includeknee, elbow, lower back, shoulder, other anatomical wraps, and apparelsuch as garments, fashion accessories, and footware.

Referring to FIG. 29 an embodiment according to the present invention ofan electromagnetic treatment apparatus integrated into a mattress pad29700 is illustrated. A mattress can also be used. Several lightweightflexible coils 29701 are integrated into the mattress pad. Thelightweight flexible coils can be constructed from fine flexibleconductive wire, conductive thread, and any other flexible conductivematerial. The flexible coils are connected to at least one end of atleast one wire 29702. However, the flexible coils can also be configuredto be directly connected to circuitry 29703 or wireless. Lightweightminiaturized circuitry 29703 that configures waveforms according to anembodiment of the present invention, is attached to at least one otherend of said at least on wire. When activated the lightweightminiaturized circuitry 29703 configures waveforms that are directed tothe flexible coils (29701) to create PEMF signals that are coupled to atarget pathway structure.

Referring to FIG. 30 an embodiment according to the present invention ofan electromagnetic treatment inductive apparatus integrated into a chestgarment 30800, such as a bra is illustrated. Several lightweightflexible coils 30801 are integrated into a bra. The lightweight flexiblecoils can be constructed from fine flexible conductive wire, conductivethread, and any other flexible conductive material. The flexible coilsare connected to at least one end of at least one wire 30802. However,the flexible coils can also be configured to be directly connected tocircuitry 30803 or wireless. Lightweight miniaturized circuitry 30803that configures waveforms according to an embodiment of the presentinvention, is attached to at least one other end of said at least onwire. When activated the lightweight miniaturized circuitry 30803configures waveforms that are directed to the flexible coils (30801) tocreate PEMF signals that are coupled to a target pathway structure.

Example 1

An embodiment according to the present invention for EMF signalconfiguration has been used on calcium dependent myosin phosphorylationin a standard enzyme assay. This enzyme pathway is known to enhance theeffects of pharmacological, chemical, cosmetic and topical agents asapplied to, upon or in human, animal and plant cells, organs, tissuesand molecules. The reaction mixture was chosen for phosphorylation rateto be linear in time for several minutes, and for sub-saturation Ca²⁺concentration. This opens the biological window for Ca²⁺/CaM to beEMF-sensitive, as happens in an injury or with the application ofpharmacological, chemical, cosmetic and topical agents as applied to,upon or in human, animal and plant cells, organs, tissues and molecules.Experiments were performed using myosin light chain (“MLC”) and myosinlight chain kinase (“MLCK”) isolated from turkey gizzard. A reactionmixture consisted of a basic solution containing 40 mM Hepes buffer, pH7.0; 0.5 mM magnesium acetate; 1 mg/ml bovine serum albumin, 0.1% (w/v)Tween 80; and 1 mM EGTA. Free Ca²⁺ was varied in the 1-7 μM range. OnceCa²⁺ buffering was established, freshly prepared 70 nM CaM, 160 nM MLCand 2 nM MLCK were added to the basic solution to form a final reactionmixture.

The reaction mixture was freshly prepared daily for each series ofexperiments and was aliquoted in 100 μL portions into 1.5 ml Eppendorftubes. All Eppendorf tubes containing reaction mixture were kept at 0°C. then transferred to a specially designed water bath maintained at37±0.1° C. by constant perfusion of water prewarmed by passage through aFisher Scientific model 900 heat exchanger. Temperature was monitoredwith a thermistor probe such as a Cole-Parmer model 8110-20, immersed inone Eppendorf tube during all experiments. Reaction was initiated with2.5 μM 32P ATP, and was stopped with Laemmli Sample Buffer solutioncontaining 30 μM EDTA. A minimum of five blank samples were counted ineach experiment. Blanks comprised a total assay mixture minus one of theactive components Ca²⁺, CaM, MLC or MLCK. Experiments for which blankcounts were higher than 300 cpm were rejected. Phosphorylation wasallowed to proceed for 5 min and was evaluated by counting ³²Pincorporated in MLC using a TM Analytic model 5303 Mark V liquidscintillation counter.

The signal comprised repetitive bursts of a high frequency waveform.Amplitude was maintained constant at 0.2 G and repetition rate was 1burst/sec for all exposures. Burst duration varied from 65 μsec to 1000μsec based upon projections of mathematical analysis of the instantinvention which showed that optimal Power SNR would be achieved as burstduration approached 500 μsec. The results are shown in FIG. 9 whereinburst width 901 in μsec is plotted on the x-axis and MyosinPhosphorylation 902 as treated/sham is plotted on the y-axis. It can beseen that the PMF effect on Ca²⁺ binding to CaM approaches its maximumat approximately 500 μsec, just as illustrated by the Power SNR model.

These results confirm that an EMF signal, configured according to anembodiment of the present invention, would maximally increase woundrepair in human, animal and plant cells, organs, tissues and moleculesfor burst durations sufficient to achieve optimal Power SNR for a givenmagnetic field amplitude.

Example 2

According to an embodiment of the present invention use of a Power SNRmodel was further verified in an in vivo wound repair model. A rat woundmodel has been well characterized both biomechanically andbiochemically, and was used in this study. Healthy, young adult maleSprague Dawley rats weighing more than 300 grams were utilized.

The animals were anesthetized with an intraperitoneal dose of Ketamine75 mg/kg and Medetomidine 0.5 mg/kg. After adequate anesthesia had beenachieved, the dorsum was shaved, prepped with a dilute betadine/alcoholsolution, and draped using sterile technique. Using a #10 scalpel, an8-cm linear incision was performed through the skin down to the fasciaon the dorsum of each rat. The wound edges were bluntly dissected tobreak any remaining dermal fibers, leaving an open wound approximately 4cm in diameter. Hemostasis was obtained with applied pressure to avoidany damage to the skin edges. The skin edges were then closed with a 4-0Ethilon running suture. Post-operatively, the animals receivedBuprenorphine 0.1-0.5 mg/kg, intraperitoneal. They were placed inindividual cages and received food and water ad libitum.

EMF exposure comprised two pulsed radio frequency waveforms. The firstwas a standard clinical PRF signal comprising a 65 μsec burst of 27.12MHz sinusoidal waves at 1 Gauss amplitude and repeating at 600bursts/sec. The second was a PRF signal reconfigured according to anembodiment of the present invention. For this signal burst duration wasincreased to 2000 μsec and the amplitude and repetition rate werereduced to 0.2 G and 5 bursts/sec respectively. PRF was applied for 30minutes twice daily.

Tensile strength was performed immediately after wound excision. Two 1cm width strips of skin were transected perpendicular to the scar fromeach sample and used to measure the tensile strength in kg/mm². Thestrips were excised from the same area in each rat to assure consistencyof measurement. The strips were then mounted on a tensiometer. Thestrips were loaded at 10 mm/min and the maximum force generated beforethe wound pulled apart was recorded. The final tensile strength forcomparison was determined by taking the average of the maximum load inkilograms per mm² of the two strips from the same wound.

The results showed average tensile strength for the 65 μsec 1 Gauss PRFsignal was 19.3±4.3 kg/mm² for the exposed group versus 13.0±3.5 kg/mm²for the control group (p<0.01), which is a 48% increase. In contrast,the average tensile strength for the 2000 μsec 0.2 Gauss PRF signal,configured according to an embodiment of the present invention using aPower SNR model was 21.2±5.6 kg/mm² for the treated group versus13.7±4.1 kg/mm² (p<0.01) for the control group, which is a 54% increase.The results for the two signals were not significantly different fromeach other.

Non-invasive, non-thermal pulsed magnetic fields are successfultherapies for healing non-union fractures, the palliative relief of painand edema and the healing of chronic wounds. The two radio frequency EMFdevices used in this study differed by burst duration, envelope,amplitude and repetition rate. That second radio frequency producednearly identical results to those produced by first radio frequencydemonstrates the validity of the EMF signal configuration according tothe present invention.

The results follow the pattern observed in clinical and basic EMFstudies. Applying correct dosimetry, that is the signal is detectable inthe EMF-sensitive pathway, the state of the target determines the degreeof effect. Thus, surrounding normal bone does not respond in aphysiologically significant manner even though it receives the same EMFdosage as cells/tissue in the fracture site. The same occurs for cellsin culture wherein a dependence upon cell cycle, state of tissue repairand the extracellular concentration of ions/ligands has been reported.Thus EMF has virtually no effect in the later stages of wound repair. Bycomparison with known biomechanical healing curve for this model, it maybe estimated that the EMF treated wounds would have reached the endstage of wound repair, approximately 1.5× faster than the sham group.

At the cellular level PMF have been shown to enhance TGF-β production.EMF of the type used for bone repair significantly increased endothelialcell tubulization and proliferation, as well as fibroblast growth factorβ-2, in vitro. Additionally, EMF signals can modulate anti-CD3 bindingat lymphocyte receptors, demonstrating EMF can reduce the inflammatoryresponse. When EMF effects occur in this cutaneous wound model,accelerated healing would be achieved, both from a reduction of time inthe inflammatory phase and subsequent acceleration of collagenproduction. The production of growth factors has been reported to beCa/CaM (calmodulin) dependent and an EMF signal has been shown toaccelerate Ca2+ binding to calmodulin. The electric field induced attissue level from the EMF signal utilized has been shown to contain theproper frequency spectrum to be detected at Ca/CaM binding pathways. Ithas also been demonstrated that inductively coupled EMF bone healingsignals can increase osteoblast proliferation in-vitro by directmodulation of Ca/CaM.

These results demonstrate that an embodiment of the present inventionallowed a EMF signal to be configured that could be produced withsignificantly lower power. The PRF signal configured according to anembodiment of the present invention, accelerated wound repair in the ratmodel in a low power manner versus that for a clinical EMF signal whichaccelerated wound repair but required more than two orders of magnitudemore power to produce.

Example 3

This study demonstrated the effect of electromagnetic fields configuredaccording an embodiment of the present invention accelerate tendonrepair in an in-vivo model.

Young adult male Sprague-Dawley rats, with a mean weight of 350 g, wereanesthetized with an intraperitoneal injection of aketamine/medetomidine 75 mg/kg/0.5 mg/kg mixture. The Achilles tendonwas disrupted and repaired. Using sterile surgical technique, a 2-cmmidline longitudinal incision was made over the right Achilles tendonwhile it was stretched by flexing the right foot. Blunt dissection wasused to separate the tendon from the surrounding tissue, which was thentransected at the middle using a scalpel. The Achilles tendon was thenimmediately repaired with 6-0 nylon suture using a modified Kesslerstitch. The plantaris tendon was divided and not repaired. The skin wassutured over the repaired tendon using interrupted 5-0 Ethilon. TheAchilles tendon was not immobilized. Postoperatively, the animalsreceived Ketoprofen for pain control.

On the first postoperative day, all animals were randomly assigned tofour treatment groups with 10 animals in each group. Randomizationfollowed the parallel group protocol wherein each animal was randomlyassigned to one treatment group until there were ten in each group.Animals remained in their assigned group. There were three active groupsthat received specific EMF treatments for two 30-min sessions per dayover a period of 3 weeks, and one identically treated sham group. TheEMF employed in this study was a pulsed radio frequency waveformcomprising a repetitive burst of 27.12 MHz sinusoidal waves emitted by aPMF-generating coil. Two configurations were employed. The first,assigned to Group 1, comprised a burst duration of 65 μsec, repeating at600 bursts/sec with an amplitude at the tendon target of 1 gauss (“G”).The second PRF waveform comprised a burst duration of 2000 μsec,repeating at 5 bursts/sec with an amplitude at the tendon target of 0.05G, assigned to Group 2, and 0.1 G, assigned to Group 3. Sham animals, nosignal, were assigned to Group 4.

The PRF signal was delivered with a single loop coil, mounted to enablea standard rat plastic cage, with all metal portions removed, to bepositioned within it. The coil was located 3.5 inches above, andhorizontal to, the floor of the cage. Five freely roaming animals weretreated with each coil. EMF signal amplitude was checked. Signalamplitude within the rat treatment cage over the normal range of ratmovement was uniform to ±10%. Signal consistency was verified weekly.There were two cages each for the sham and active groups, and each cagehad its individual coded EMF exposure system. EMF treatment was carriedout twice daily for 30-min sessions until sacrifice. Sham animals weretreated in identical cages equipped with identical coils.

At the end of the 3-week treatment period, the Achilles tendon washarvested by proximally severing the muscle bellies arising from thetendon and distally disarticulating the ankle, keeping the calcaneousand foot attached. All extraneous soft and hard tissues were removedfrom the calcaneous-Achilles tendon complex. Tensile strength testingwas done immediately after harvest. The tendon, in continuity with thecalcaneal bone, was fixed between two metal clamps so as to maintain aphysiologically appropriate foot dorsiflexion, compared to thevertically oriented Achilles tendon. The tendons were then pulled apartat a constant speed of 0.45 mm/sec until failure, and the peak tensilestrength was recorded. All analyzable tendons failed at the originaltransection. The tensile strengths from a total of 38 tendons wereavailable for analysis.

Mean tensile strength was compared for each group at 3 weeks post tendontransection and data were analyzed. Tensile strength was calculated asthe maximum breaking strength in kilograms per cross-sectional area insquare centimeters. Tendons treated with the 65 μsec signal in Group 1had a mean breaking strength of 99.4±14.6 kg/cm2 compared to 80.6±16.6kg/cm2 for the sham-treated group in Group 4. This represented a 24%increase in breaking strength vs. the sham group at 21 days, which wasnot statistically significant (p=0.055). Tendons from Groups 2 and 3,treated with the 2000 μsec signals, had significantly higher meanbreaking strengths of 129.4±27.8 kg/cm2 and 136.4±31.6 kg/cm2 for the0.05 G and 0.1 G signals, respectively, vs. the sham exposure group80.6±16.6 kg/cm2. The mean strengths for both Groups 2 and 3 were 60%and 69% higher, respectively, at the end of 3 weeks of treatment,compared to the sham group. This increase in strength was statisticallysignificant (p<0.001); however, the difference in mean tensile strengthbetween Groups 2 and 3 was not statistically significant (p=0.541). Thedifferences in mean tensile strength between Group 1 (65 μsec burst) andGroups 2 and 3 (2000 μsec burst) was statistically significant (p<0.05).

The results presented here demonstrate that non-invasive pulsedelectromagnetic fields can produce up to a 69% increase in rat Achillestendon breaking strength vs. sham-treated tendons at 21 days posttransection. All signals utilized in this study accelerated tendonrepair, however greatest acceleration was obtained with waveformsconfigured according to a transduction mechanism involving Ca2+ binding.

In a manner similar to bone and wound repair, tendon repair for bothepitenon and synovial-sheathed tendons begins with an inflammatory stagethat generally involves infiltration of inflammatory cells such asmacrophages, neutrophils, and T-lymphocytes. This is followed byangiogenesis, fibroblast proliferation, and collagen mainly type III,production. Finally, cells and collagen fibrils orient to achievemaximum mechanical strength. These phases all occur in bone and woundrepair, in which EMF has demonstrated effects, particularly ininflammatory, angiogenesis, and cell proliferation stages.

An EMF transduction pathway involves ion binding in regulatory pathwaysinvolving growth factor release. Production of many of the growthfactors and cytokines involved in tissue growth and repair is dependenton Ca/CaM calmodulin. EMF has been shown to accelerate Ca2+ binding tocalmodulin. The 0.05 and 0.1 G signals utilized in this study wereconfigured using a Ca/CaM transduction pathway. The objective was toproduce sufficient electric field amplitude that is dose, within thefrequency response of Ca2+ binding. This would result in a lower power,more effective signal. The model demonstrated that microsecond rangeburst durations satisfy these objectives at amplitudes in the 0.05 Grange. The 0.1 G signal was added to assure that the small size of therat tendon target did not limit the induced current pathway and reducethe expected dose.

EMF accelerates bone repair by accelerating return to intact breakingstrength. The sham-treated fractures eventually reach the samebiomechanical end point, but with increased morbidity. Biomechanicalacceleration in a linear full-thickness cutaneous wound in the rat wasobserved. EMF accelerated wound repair by approximately 60% at 21 days,with intact breaking strength achieved about 50% sooner than theuntreated wounds.

Having described embodiments for an apparatus and a method for enhancingpharmacological effects, it is noted that modifications and variationscan be made by persons skilled in the art in light of the aboveteachings. It is therefore to be understood that changes may be made inthe particular embodiments of the invention disclosed which are withinthe scope and spirit of the invention as defined by the appended claims.

Part 5

An embodiment according to the present invention provides a higherspectral density to a pulse burst envelope resulting in enhancedeffectiveness of therapy upon relevant dielectric pathways, such as,cellular membrane receptors, ion binding to cellular enzymes and generaltransmembrane potential changes. An embodiment according to the presentinvention increases the number of frequency components transmitted torelevant cellular pathways, thereby providing access to a larger rangeof biophysical phenomena applicable to known healing mechanisms, forexample modulation of growth factor and cytokine release, and ionbinding at regulatory molecules. By applying a random, or other highspectral density envelope, according to a mathematical model defined bySNR or Power SNR in a transduction pathway, to a pulse burst envelope ofmono- or bi-polar rectangular or sinusoidal pulses inducing peakelectric fields between 10-6 and 10 volts percentimeter (V/cm), agreater effect could be accomplished on biological healing processesapplicable to both soft and hard tissues thereby enhancing effectivenessof pharmacological, chemical, cosmetic and topical agents.

An advantageous result of the present invention, is that by applying ahigh spectral density voltage envelope as the modulating or pulse-burstdefining parameter, according to a mathematical model defined by SNR orPower SNR in a transduction pathway, the power requirement for suchamplitude modulated pulse bursts can be significantly lower than that ofan unmodulated pulse burst containing pulses within the same frequencyrange. Accordingly, the advantages of enhanced transmitted dosimetry tothe relevant dielectric target pathways and of decreased powerrequirement are achieved.

An additional advantage of the present invention relates to enhancedeffectiveness of pharmacological, chemical, cosmetic and topical agentsas applied to, upon or on human, animal and plant cells, organs, tissuesand molecules by accelerating the agents intended effects and improvingefficacy.

Induced time-varying currents from PEMF or PRF devices flow in a targetpathway structure such as a molecule, cell, tissue, and organ, and it isthese currents that are a stimulus to which cells and tissues can reactin a physiologically meaningful manner. The electrical properties of atarget pathway structure affect levels and distributions of inducedcurrent. Molecules, cells, tissue, and organs are all in an inducedcurrent pathway such as cells in a gap junction contact. Ion or ligandinteractions at binding sites on macromolecules that may reside on amembrane surface are voltage dependent processes, for exampleelectrochemical, that can respond to an induced electromagnetic field(“E”). Induced current arrives at these sites via a surrounding ionicmedium. The presence of cells in a current pathway causes an inducedcurrent (“J”) to decay more rapidly with time (“J(t)”). This is due toan added electrical impedance of cells from membrane capacitance andtime constants of binding and other voltage sensitive membrane processessuch as membrane transport.

Equivalent electrical circuit models representing various membrane andcharged interface configurations have been derived. For example, inCalcium (“Ca2+”) binding, the change in concentration of bound Ca2+ at abinding site due to induced E may be described in a frequency domain byan impedance expression such as:

${Z_{b}(\omega)} = {R_{ion} + \frac{1}{i\; \omega \; C_{ion}}}$

which has the form of a series resistance-capacitance electricalequivalent circuit. Where ω is angular frequency defined as 2πf, where fis frequency, i=−1^(1/2), Z_(b)(ω) is the binding impedance, and R_(ion)and C_(ion) are equivalent binding resistance and capacitance of an ionbinding pathway. The value of the equivalent binding time constant,τ_(ion)=R_(ion)C_(ion), is related to a ion binding rate constant,k_(b), via τ_(ion)=R_(ion)C_(ion)=1/k_(b). Thus, the characteristic timeconstant of this pathway is determined by ion binding kinetics.

Induced E from a PEMF or PRF signal can cause current to flow into anion binding pathway and affect the number of Ca²⁺ ions bound per unittime. An electrical equivalent of this is a change in voltage across theequivalent binding capacitance C_(ion), which is a direct measure of thechange in electrical charge stored by C_(ion). Electrical charge isdirectly proportional to a surface concentration of Ca²⁺ ions in thebinding site, that is storage of charge is equivalent to storage of ionsor other charged species on cell surfaces and junctions. Electricalimpedance measurements, as well as direct kinetic analyses of bindingrate constants, provide values for time constants necessary forconfiguration of a PMF waveform to match a bandpass of target pathwaystructures. This allows for a required range of frequencies for anygiven induced E waveform for optimal coupling to target impedance, suchas bandpass.

Ion binding to regulatory molecules is a frequent EMF target, forexample Ca²⁺ binding to calmodulin (“CaM”). Use of this pathway is basedupon acceleration of wound repair, for example bone repair, thatinvolves modulation of growth factors released in various stages ofrepair. Growth factors such as platelet derived growth factor (“PDGF”),fibroblast growth factor (“FGF”), and epidermal growth factor (“EGF”)are all involved at an appropriate stage of healing. Angiogenesis isalso integral to wound repair and modulated by PMF. All of these factorsare Ca/CaM-dependent.

Utilizing a Ca/CaM pathway a waveform can be configured for whichinduced power is sufficiently above background thermal noise power.Under correct physiological conditions, this waveform can have aphysiologically significant bioeffect.

Application of a Power SNR model to Ca/CaM requires knowledge ofelectrical equivalents of Ca²⁺ binding kinetics at CaM. Within firstorder binding kinetics, changes in concentration of bound Ca²⁺ at CaMbinding sites over time may be characterized in a frequency domain by anequivalent binding time constant, τ_(ion)=R_(ion)C_(ion), where R_(ion)and C_(ion) are equivalent binding resistance and capacitance of the ionbinding pathway. τ_(ion) is related to a ion binding rate constant,k_(b), via τ_(ion)=R_(ion)C_(ion)=1/k_(b). Published values for k_(b)can then be employed in a cell array model to evaluate SNR by comparingvoltage induced by a PRF signal to thermal fluctuations in voltage at aCaM binding site. Employing numerical values for PMF response, such asV_(max)=6.5×10⁻⁷ sec⁻¹, =2.50/1, K_(D)=30 μM, [Ca²⁺CaM]=K_(D)(+[CaM]),yields k_(b)=665 sec⁻¹ (τ_(ion)=1.5 msec). Such a value for τ_(ion) canbe employed in an electrical equivalent circuit for ion binding whilepower SNR analysis can be performed for any waveform structure.

According to an embodiment of the present invention a mathematical modelcan be configured to assimilate that thermal noise is present in allvoltage dependent processes and represents a minimum thresholdrequirement to establish adequate SNR. Power spectral density, S_(n)(ω),of thermal noise can be expressed as:

S _(n)(ω)=4kT Re[Z _(M)(x,ω)]

where Z_(M)(x, ω) is electrical impedance of a target pathway structure,x is a dimension of a target pathway structure and Re denotes a realpart of impedance of a target pathway structure. Z_(M)(x, ω) can beexpressed as:

${Z_{M}( {x,\omega} )} = {\lbrack \frac{R_{e} + R_{i} + R_{g}}{\gamma} \rbrack {\tanh ( {\gamma \; x} )}}$

This equation clearly shows that electrical impedance of the targetpathway structure, and contributions from extracellular fluid resistance(“R_(e)”), intracellular fluid resistance (“R_(i)”) and intermembraneresistance (“R_(g)”) which are electrically connected to a targetpathway structure, all contribute to noise filtering.

A typical approach to evaluation of SNR uses a single value of a rootmean square (RMS) noise voltage. This is calculated by taking a squareroot of an integration of S_(n)(ω)=4 kT Re [Z_(M)(x, ω)] over allfrequencies relevant to either complete membrane response, or tobandwidth of a target pathway structure. SNR can be expressed by aratio:

${SNR} = \frac{{V_{M}(\omega)}}{RMS}$

where |V_(M)(ω)| is maximum amplitude of voltage at each frequency asdelivered by a chosen waveform to the target pathway structure.

An embodiment according to the present invention comprises a pulse burstenvelope having a high spectral density, so that the effect of therapyupon the relevant dielectric pathways, such as, cellular membranereceptors, ion binding to cellular enzymes and general transmembranepotential changes, is enhanced. Accordingly by increasing a number offrequency components transmitted to relevant cellular pathways, a largerange of biophysical phenomena, such as modulating growth factor andcytokine release and ion binding at regulatory molecules, applicable toknown tissue growth mechanisms is accessible. According to an embodimentof the present invention applying a random, or other high spectraldensity envelope, to a pulse burst envelope of mono- or bi-polarrectangular or sinusoidal pulses inducing peak electric fields betweenabout 10⁻⁸ and about 100 V/cm, produces a greater effect on biologicalhealing processes applicable to both soft and hard tissues.

According to yet another embodiment of the present invention by applyinga high spectral density voltage envelope as a modulating or pulse-burstdefining parameter, power requirements for such amplitude modulatedpulse bursts can be significantly lower than that of an unmodulatedpulse burst containing pulses within a similar frequency range. This isdue to a substantial reduction in duty cycle within repetitive bursttrains brought about by imposition of an irregular, and preferablyrandom, amplitude onto what would otherwise be a substantially uniformpulse burst envelope. Accordingly, the dual advantages, of enhancedtransmitted dosimetry to the relevant dielectric pathways and ofdecreased power requirement are achieved.

Referring to FIG. 32, wherein FIG. 32 is a flow diagram of a methodaccording to an embodiment of the present invention, for enhancingeffectiveness of pharmacological, chemical, cosmetic and topical agentsused to treat stem cells, tissues, cells, organs, and molecules bydelivering electromagnetic signals that can be pulsed, to target pathwaystructures such as ions and ligands of animals and humans, fortherapeutic and prophylactic purposes. Target pathway structures canalso include but are not limited to stem cells, tissues, cells, organs,and molecules. Enhancing effectiveness of pharmacological, chemical,cosmetic and topical agents includes but is not limited to increasedabsorption rate, decreased effective dosages, faster delivery rates atan organism level; and increased binding kinetics and transport kineticslevel at a molecular and cellular level. At least one reactive agent isapplied to a target pathway structure (Step 32101). Reactive agentsinclude but are not limited to pharmacological agents, chemical agents,cosmetic agents, topical agents, and genetic agents. Reactive agents canbe ingested, applied topically, applied intravenously, intramuscularly,or by any other manner known within the medical community that causesinteraction of substances with a target pathway structure, such asiontophoresis, X and light radiation, and heat. Pharmacological agentsinclude but are not limited to antibiotics, growth factors,chemotherapeutic agents, antihistamines, Angiotensin inhibitors, betablockers, statins, and anti-inflammatory drugs. Chemical agents includebut are not limited to hydrogen peroxide, betadine, and alcohol. Topicalagents include but are not limited to antibiotics, creams, retinol,benzoyl peroxide, tolnaftate, menthol, emollients, oils, lanolin,squalene, aloe vera, anti-oxidants, fatty acid, fatty acid ester, codliver oil, alpha-tocopherol, petroleum, hydrogenated polybutene, vitaminA, vitamin E, topical proteins, and collagens. Cosmetic agents includebut are not limited to make-up, eye-liner, and blush. Genetic agentsinclude but are not limited to genes, DNA, and chromosomes.

Configuring at least one waveform having at least one waveform parameterto be coupled to the target pathway structure such as ions and ligands(Step 32102).

The at least one waveform parameter is selected to maximize at least oneof a signal to noise ratio and a Power Signal to Noise ratio in a targetpathway structure so that a waveform is detectable in the target pathwaystructure above its background activity (Step 32102) such as baselinethermal fluctuations in voltage and electrical impedance at a targetpathway structure that depend upon a state of a cell and tissue, that iswhether the state is at least one of resting, growing, replacing, andresponding to injury to produce physiologically beneficial results. Tobe detectable in the target pathway structure the value of said at leastone waveform parameter is chosen by using a constant of said targetpathway structure to evaluate at least one of a signal to noise ratio,and a Power signal to noise ratio, to compare voltage induced by said atleast one waveform in said target pathway structure to baseline thermalfluctuations in voltage and electrical impedance in said target pathwaystructure whereby bioeffective modulation occurs in said target pathwaystructure by said at least one waveform by maximizing said at least oneof signal to noise ratio and Power signal to noise ratio, within abandpass of said target pathway structure.

A preferred embodiment of a generated electromagnetic signal iscomprised of a burst of arbitrary waveforms having at least one waveformparameter that includes a plurality of frequency components ranging fromabout 0.01 Hz to about 100 MHz wherein the plurality of frequencycomponents satisfies a Power SNR model (Step 32103). A repetitiveelectromagnetic signal can be generated for example inductively orcapacitively, from said configured at least one waveform (Step 32104).The electromagnetic signal can also be non-repetitive. Theelectromagnetic signal is coupled to a target pathway structure such asions and ligands by output of a coupling device such as an electrode oran inductor, placed in close proximity to the target pathway structure(Step 32105). Coupling of the electromagnetic signal to a target pathwaystructure can occur adjunctively, for example at any time prior toapplying a reactive agent, at the same time a reactive agent is beingapplied, or after the time a reactive agent has been applied. Thecoupling enhances blood flow and modulation of binding of ions andligands to regulatory molecules in molecules, tissues, cells, and organsthereby enhancing the reactive agents' bioeffectiveness.

FIG. 33 illustrates a preferred embodiment of an apparatus according tothe present invention. The apparatus is self-contained, lightweight, andportable. A miniature control circuit 33201 is coupled to an end of atleast one connector 33202 such as wire however the control circuit canalso operate wirelessly. The opposite end of the at least one connectoris coupled to a generating device such as an electrical coil 33203. Theminiature control circuit 33201 is constructed in a manner that appliesa mathematical model that is used to configure waveforms. The configuredwaveforms have to satisfy Power SNR so that for a given and known targetpathway structure, it is possible to choose waveform parameters thatsatisfy Power SNR so that a waveform produces physiologically beneficialresults, for example bioeffective modulation, and is detectable in thetarget pathway structure above its background activity. A preferredembodiment according to the present invention applies a mathematicalmodel to induce a time-varying magnetic field and a time-varyingelectric field in a target pathway structure such as ions and ligands,comprising about 0.1 to about 100 msec bursts of about 1 to about 100microsecond rectangular pulses repeating at about 0.1 to about 100pulses per second. Peak amplitude of the induced electric field isbetween about 1 uV/cm and about 100 mV/cm, varied according to amodified 1/f function where f=frequency. A waveform configured using apreferred embodiment according to the present invention may be appliedto a target pathway structure such as ions and ligands for a preferredtotal exposure time of under 1 minute to 240 minutes daily. Howeverother exposure times can be used. Waveforms configured by the miniaturecontrol circuit 33201 are directed to a generating device 33203 such aselectrical coils via connector 33202. The generating device 33203delivers a pulsing magnetic field that can be used to provide treatmentto a target pathway structure such as tissue. The miniature controlcircuit applies a pulsing magnetic field for a prescribed time and canautomatically repeat applying the pulsing magnetic field for as manyapplications as are needed in a given time period, for example 10 timesa day. The miniature control circuit can be configured to beprogrammable applying pulsing magnetic fields for any time repetitionsequence. A preferred embodiment according to the present invention canenhance the pharmacological, chemical, cosmetic and topical agents'effectiveness by being incorporated into a positioning device 33204, forexample a bed. Coupling a pulsing magnetic field to a target pathwaystructure such as ions and ligands, therapeutically and prophylacticallyreduces inflammation thereby advantageously reducing pain, promotinghealing in targeted areas, and enhancing interactions ofpharmacological, chemical, cosmetic and topical agents with a targetpathway structure. When electrical coils are used as the generatingdevice 33203, the electrical coils can be powered with a time varyingmagnetic field that induces a time varying electric field in a targetpathway structure according to Faraday's law. An electromagnetic signalgenerated by the generating device 33203 can also be applied usingelectrochemical coupling, wherein electrodes are in direct contact withskin or another outer electrically conductive boundary of a targetpathway structure. Yet in another embodiment according to the presentinvention, the electromagnetic signal generated by the generating device33203 can also be applied using electrostatic coupling wherein an airgap exists between a generating device 33203 such as an electrode and atarget pathway structure such as ions and ligands. An advantage of thepreferred embodiment according to the present invention is that itsultra lightweight coils and miniaturized circuitry allow for use withcommon physical therapy treatment modalities and at any for whichgrowth, pain relief, and tissue and organ healing is desired. Anadvantageous result of application of the preferred embodiment accordingto the present invention is that tissue growth, repair, and maintenancecan be accomplished and enhanced anywhere and at anytime, for examplewhile driving a car or watching television. Yet another advantageousresult of application of the preferred embodiment is that growth,repair, and maintenance of molecules, cells, tissues, and organs can beaccomplished and enhanced anywhere and at anytime, for example whiledriving a car or watching television.

FIG. 34 depicts a block diagram of a preferred embodiment according tothe present invention of a miniature control circuit 34300. Theminiature control circuit 34300 produces waveforms that drive agenerating device such as wire coils described above in FIG. 33. Theminiature control circuit can be activated by any activation means suchas an on/off switch. The miniature control circuit 34300 has a powersource such as a lithium battery 34301. A preferred embodiment of thepower source has an output voltage of 3.3 V but other voltages can beused. In another embodiment according to the present invention the powersource can be an external power source such as an electric currentoutlet such as an AC/DC outlet, coupled to the present invention forexample by a plug and wire. A switching power supply 34302 controlsvoltage to a micro-controller 34303. A preferred embodiment of themicro-controller 34303 uses an 8 bit 4 MHz micro-controller 34303 butother bit MHz combination micro-controllers may be used. The switchingpower supply 34302 also delivers current to storage capacitors 34304. Apreferred embodiment of the present invention uses storage capacitorshaving a 220 uF output but other outputs can be used. The storagecapacitors 34304 allow high frequency pulses to be delivered to acoupling device such as inductors (Not Shown). The micro-controller34303 also controls a pulse shaper 34305 and a pulse phase timingcontrol 34306. The pulse shaper 34305 and pulse phase timing control34306 determine pulse shape, burst width, burst envelope shape, andburst repetition rate. An integral waveform generator, such as a sinewave or arbitrary number generator can also be incorporated to providespecific waveforms. A voltage level conversion sub-circuit 34307controls an induced field delivered to a target pathway structure. Aswitching Hexfet 34308 allows pulses of randomized amplitude to bedelivered to output 34309 that routes a waveform to at least onecoupling device such as an inductor. The micro-controller 34303 can alsocontrol total exposure time of a single treatment of a target pathwaystructure such as a molecule, cell, tissue, and organ. The miniaturecontrol circuit 34300 can be constructed to be programmable and apply apulsing magnetic field for a prescribed time and to automatically repeatapplying the pulsing magnetic field for as many applications as areneeded in a given time period, for example 10 times a day. A preferredembodiment according to the present invention uses treatments times ofabout 10 minutes to about 30 minutes.

Referring to FIGS. 35A and 35B a preferred embodiment according to thepresent invention of a coupling device 35400 such as an inductor isshown. The coupling device 35400 can be an electric coil 35401 woundwith single or multistrand flexible wire 35402 however solid wire canalso be used. In a preferred embodiment according to the presentinvention the wire is made of copper but other materials can be used.The multistrand flexible magnetic wire 35402 enables the electric coil35401 to conform to specific anatomical configurations such as a limb orjoint of a human or animal. A preferred embodiment of the electric coil35401 comprises about 1 to about 1000 turns of about 0.01 mm to about0.1 mm diameter at least one of single magnet wire and multistrandmagnet wire, wound on an initially circular form having an outerdiameter between about 2.5 cm and about 50 cm but other numbers of turnsand wire diameters can be used. A preferred embodiment of the electriccoil 401 can be encased with a non-toxic PVC mould 35403 but othernon-toxic moulds can also be used. The electric coil can also beincorporated in dressings, bandages, garments, and other structurestypically used for wound treatment.

Referring to FIG. 36 an embodiment according to the present invention ofa waveform 36500 is illustrated. A pulse 36501 is repeated within aburst 36502 that has a finite duration 36503. The duration 36503 is suchthat a duty cycle which can be defined as a ratio of burst duration tosignal period is between about 1 to about 10⁻⁵. A preferred embodimentaccording to the present invention utilizes pseudo rectangular 10microsecond pulses for pulse 36501 applied in a burst 36502 for about 10to about 50 msec having a modified 1/f amplitude envelope 36504 and witha finite duration 36503 corresponding to a burst period of between about0.1 and about 10 seconds, but other waveforms, envelopes, and burstperiods that follow a mathematical model such as SNR and Power SNR, maybe used.

FIG. 37 illustrates a preferred embodiment according to the presentinvention of a positioning device such as a wrist support. A positioningdevice 37600 such as a wrist support 37601 is worn on a human wrist37602. The positioning device can be constructed to be portable, can beconstructed to be disposable, and can be constructed to be implantable.The positioning device can be used in combination with the presentinvention in a plurality of ways, for example incorporating the presentinvention into the positioning device for example by stitching, affixingthe present invention onto the positioning device for example byVelcro®, and holding the present invention in place by constructing thepositioning device to be elastic.

In another embodiment according to the present invention, the presentinvention can be constructed as a stand-alone device of any size with orwithout a positioning device, to be used anywhere for example at home,at a clinic, at a treatment center, and outdoors. The wrist support 601can be made with any anatomical and support material, such as neoprene.Coils 37603 are integrated into the wrist support 37601 such that asignal configured according to the present invention, for example thewaveform depicted in FIG. 36, is applied from a dorsal portion that is,the top of the wrist to a plantar portion that is the bottom of thewrist. Micro-circuitry 37604 is attached to the exterior of the wristsupport 37601 using a fastening device such as Velcro®. (Not Shown). Themicro-circuitry is coupled to one end of at least one connecting devicesuch as a flexible wire 37605. The other end of the at least oneconnecting device is coupled to the coils 37603. Other embodimentsaccording to the present invention of the positioning device includeknee, elbow, lower back, shoulder, other anatomical wraps, and apparelsuch as garments, fashion accessories, and footware.

Referring to FIG. 38 an embodiment according to the present invention ofan electromagnetic treatment apparatus integrated into a mattress pad38700 is illustrated. A mattress can also be used. Several lightweightflexible coils 38701 are integrated into the mattress pad. Thelightweight flexible coils can be constructed from fine flexibleconductive wire, conductive thread, and any other flexible conductivematerial. The flexible coils are connected to at least one end of atleast one wire 38702. However, the flexible coils can also be configuredto be directly connected to circuitry 38703 or wireless. Lightweightminiaturized circuitry 38703 that configures waveforms according to anembodiment of the present invention, is attached to at least one otherend of said at least on wire. When activated the lightweightminiaturized circuitry 38703 configures waveforms that are directed tothe flexible coils (38701) to create PEMF signals that are coupled to atarget pathway structure.

Example 1

An embodiment according to the present invention for EMF signalconfiguration has been used on calcium dependent myosin phosphorylationin a standard enzyme assay. This enzyme pathway is known to enhance theeffects of pharmacological, chemical, cosmetic and topical agents asapplied to, upon or in human, animal and plant cells, organs, tissuesand molecules. The reaction mixture was chosen for phosphorylation rateto be linear in time for several minutes, and for sub-saturation Ca²⁺concentration. This opens the biological window for Ca²⁺/CaM to beEMF-sensitive, as happens in an injury or with the application ofpharmacological, chemical, cosmetic and topical agents as applied to,upon or in human, animal and plant cells, organs, tissues and molecules.Experiments were performed using myosin light chain (“MLC”) and myosinlight chain kinase (“MLCK”) isolated from turkey gizzard. A reactionmixture consisted of a basic solution containing 40 mM Hepes buffer, pH7.0; 0.5 mM magnesium acetate; 1 mg/ml bovine serum albumin, 0.1% (w/v)Tween 80; and 1 mM EGTA. Free Ca²⁺ was varied in the 1-7 μM range. OnceCa²⁺ buffering was established, freshly prepared 70 nM CaM, 160 nM MLCand 2 nM MLCK were added to the basic solution to form a final reactionmixture.

The reaction mixture was freshly prepared daily for each series ofexperiments and was aliquoted in 100 μL portions into 1.5 ml Eppendorftubes. All Eppendorf tubes containing reaction mixture were kept at 0°C. then transferred to a specially designed water bath maintained at37±0.1° C. by constant perfusion of water prewarmed by passage through aFisher Scientific model 900 heat exchanger. Temperature was monitoredwith a thermistor probe such as a Cole-Parmer model 8110-20, immersed inone Eppendorf tube during all experiments. Reaction was initiated with2.5 μM 32P ATP, and was stopped with Laemmli Sample Buffer solutioncontaining 30 μM EDTA. A minimum of five-blank samples were counted ineach experiment. Blanks comprised a total assay mixture minus one of theactive components Ca²⁺, CaM, MLC or MLCK. Experiments for which blankcounts were higher than 300 cpm were rejected. Phosphorylation wasallowed to proceed for 5 min and was evaluated by counting 32pincorporated in MLC using a TM Analytic model 5303 Mark V liquidscintillation counter.

The signal comprised repetitive bursts of a high frequency waveform.Amplitude was maintained constant at 0.2 G and repetition rate was 1burst/sec for all exposures. Burst duration varied from 65 μsec to 1000μsec based upon projections of mathematical analysis of the instantinvention which showed that optimal Power SNR would be achieved as burstduration approached 500 μsec. The results are shown in FIG. 39 whereinburst width 39801 in μsec is plotted on the x-axis and MyosinPhosphorylation 39802 as treated/sham is plotted on the y-axis. It canbe seen that the PMF effect on Ca²⁺ binding to CaM approaches itsmaximum at approximately 500 μsec, just as illustrated by the Power SNRmodel.

These results confirm that an EMF signal, configured according to anembodiment of the present invention, would maximally increase the effectof pharmacological, chemical, cosmetic and topical agents as applied to,upon or in human, animal and plant cells, organs, tissues and moleculesfor burst durations sufficient to achieve optimal Power SNR for a givenmagnetic field amplitude.

Example 2

This study determined to what extent treatment with pulsedelectromagnetic frequency (“PEMF”) waveforms affects blood perfusion ina treated region. All testing was done in a temperature controlled room(23 to 24° C.) with the subject seated on a comfortable easy chair. Oneach arm a non-metallic laser Doppler probe was affixed withdouble-sided tape to a medial forearm site approximately 5 cm distal tothe antecubital space. A temperature sensing thermistor for surfacetemperature measurements was placed approximately 1 cm distal to theouter edge of the probes and secured with tape. A towel was draped overeach forearm to diminish the direct effects of any circulating aircurrents. With the subject resting comfortably, the skin temperature ofeach arm was monitored. During this monitoring interval the excitationcoil for producing the PEMF waveform according to the instant inventionwas positioned directly above the Laser Doppler probe of the rightforearm at a vertical distance of approximately 2 cm from the skinsurface. When the monitored skin temperature reached a steady statevalue, the data acquisition phase was begun. This consisted of a 20minute baseline interval followed by a 45 minute interval in which thePEMF waveform was applied.

Skin temperature was recorded at five minute intervals during the entireprotocol. Blood perfusion signals as determined with the Laser DopplerFlowmeter (“LDF”) were continuously displayed on a chart recorder andsimultaneously acquired by a computer following analog to digitalconversion. The LDF signals were time averaged by the computer duringeach contiguous five minute interval of measurement to produce a singleaveraged perfusion value for each interval. At the end of the procedurethe relative magnetic field strength at the skin site was measured witha 1 cm diameter loop which was coupled to a specially designed andcalibrated metering system.

For each subject the baseline perfusion for the treated arm and thecontrol arm was determined as the average during the 20 minute baselineinterval. Subsequent perfusion values, following the start of PEMFtreatment, was expressed as a percentage of this baseline. Comparisonbetween the treated and control arms were done using analysis ofvariance with arm (treated vs. control) as the grouping variables andwith time as a repeated measure.

FIG. 40 summarizes the time course of the perfusion change found duringtreatment for the nine subjects studied with time being plotted on thex-axis 40901 and perfusion on the y-axis 40902. Analysis showssignificant treatment-time interaction (p=0.03) with a significantly(p<0.01) elevated blood perfusion in the treated arm after 40 minutes ofPEMF treatment. The absolute values of baseline perfusion (mv) did notdiffer between control and treated arms. Analysis of covariance with thebaseline perfusion in absolute units (mv) as the covariate also shows anoverall difference between treated and control arms (p<0.01).

A main finding of the present investigational study is that PEMFtreatment, when applied in the manner described, is associated with asignificant augmentation in their resting forearm skin microvascularperfusion. This augmentation, which averages about 30% as compared withresting pre-treatment levels, occurs after about 40 minutes of treatmentwhereas no such augmentation is evident in the contralateral non-treatedarm. This allows the increased flow of pharmacological, chemical,topical, cosmetic, and genetic agents to the intended tissue target.

Having described embodiments for an apparatus and a method for enhancingpharmacological effects, it is noted that modifications and variationscan be made by persons skilled in the art in light of the aboveteachings. It is therefore to be understood that changes may be made inthe particular embodiments of the invention disclosed which are withinthe scope and spirit of the invention as defined by the appended claims.

Part 6

Induced time-varying currents from PEMF or PRF devices flow in a targetpathway structure such as a molecule, cell, tissue, and organ, and it isthese currents that are a stimulus to which cells and tissues can reactin a physiologically meaningful manner. The electrical properties of atarget pathway structure affect levels and distributions of inducedcurrent. Molecules, cells, tissue, and organs are all in an inducedcurrent pathway such as cells in a gap junction contact. Ion or ligandinteractions at binding sites on macromolecules that may reside on amembrane surface are voltage dependent processes, that iselectrochemical, that can respond to an induced electromagnetic field(“E”). Induced current arrives at these sites via a surrounding ionicmedium. The presence of cells in a current pathway causes an inducedcurrent (“J”) to decay more rapidly with time (“J(t)”). This is due toan added electrical impedance of cells from membrane capacitance andtime constants of binding and other voltage sensitive membrane processessuch as membrane transport.

Equivalent electrical circuit models representing various membrane andcharged interface configurations have been derived. For example, inCalcium (“Ca²⁺”) binding, the change in concentration of bound Ca²⁺ at abinding site due to induced E may be described in a frequency domain byan impedance expression such as:

${Z_{b}(\omega)} = {R_{ion} + \frac{1}{i\; \omega \; C_{ion}}}$

which has the form of a series resistance-capacitance electricalequivalent circuit. Where ω is angular frequency defined as 2πf, where fis frequency, i=−1^(1/2), Z_(b)(ω) is the binding impedance, and R_(ion)and C_(ion) are equivalent binding resistance and capacitance of an ionbinding pathway. The value of the equivalent binding time constant,τ_(ion)=R_(ion)C_(ion), is related to a ion binding rate constant,k_(b), via τ_(ion)=R_(ion)C_(ion)=1/k_(b). Thus, the characteristic timeconstant of this pathway is determined by ion binding kinetics.

Induced E from a PEMF or PRF signal can cause current to flow into anion binding pathway and affect the number of Ca²⁺ ions bound per unittime. An electrical equivalent of this is a change in voltage across theequivalent binding capacitance C_(ion), which is a direct measure of thechange in electrical charge stored by C_(ion). Electrical charge isdirectly proportional to a surface concentration of Ca²⁺ ions in thebinding site, that is storage of charge is equivalent to storage of ionsor other charged species on cell surfaces and junctions. Electricalimpedance measurements, as well as direct kinetic analyses of bindingrate constants, provide values for time constants necessary forconfiguration of a PMF waveform to match a bandpass of target pathwaystructures. This allows for a required range of frequencies for anygiven induced E waveform for optimal coupling to target impedance, suchas bandpass.

Ion binding to regulatory molecules is a frequent EMF target, forexample Ca²⁺ binding to calmodulin (“CaM”). Use of this pathway is basedupon acceleration of tissue repair, for example bone repair, woundrepair, hair repair, and repair of other molecules, cells, tissues, andorgans that involves modulation of growth factors released in variousstages of repair. Growth factors such as platelet derived growth factor(“PDGF”), fibroblast growth factor (“FGF”), and epidermal growth factor(“EGF”) are all involved at an appropriate stage of healing.Angiogenesis and neovascularization are also integral to tissue growthand repair and can be modulated by PMF. All of these factors areCa/CaM-dependent.

Utilizing a Ca/CaM pathway a waveform can be configured for whichinduced power is sufficiently above background thermal noise power.Under correct physiological conditions, this waveform can have aphysiologically significant bioeffect.

Application of a Power SNR model to Ca/CaM requires knowledge ofelectrical equivalents of Ca²⁺ binding kinetics at CaM. Within firstorder binding kinetics, changes in concentration of bound Ca²⁺ at CaMbinding sites over time may be characterized in a frequency domain by anequivalent binding time constant, τ_(ion)=R_(ion)C_(ion), where R_(ion)and C_(ion) are equivalent binding resistance and capacitance of the ionbinding pathway. τ_(ion) is related to a ion binding rate constant,k_(b), via τ_(ion)=R_(ion)C_(ion)=1/k_(b). Published values for k_(b)can then be employed in a cell array model to evaluate SNR by comparingvoltage induced by a PRF signal to thermal fluctuations in voltage at aCaM binding site. Employing numerical values for PMF response, such asV_(max)=6.5×10⁻⁷ sec⁻¹, =2.50/1, K_(D)=30 μM, [Ca²⁺CaM]=K_(D)(+[CaM]),yields k_(b)=665 sec⁻¹ (τ_(ion)=1.5 msec). Such a value for τ_(ion) canbe employed in an electrical equivalent circuit for ion binding whilepower SNR analysis can be performed for any waveform structure.

According to an embodiment of the present invention a mathematical modelcan be configured to assimilate that thermal noise is present in allvoltage dependent processes and represents a minimum thresholdrequirement to establish adequate SNR. Power spectral density, S_(n)(ω),of thermal noise can be expressed as:

S _(n)(ω)=4kT Re[Z _(M)(x,ω)]

where Z_(M)(x,ω) is electrical impedance of a target pathway structure,x is a dimension of a target pathway structure and Re denotes a realpart of impedance of a target pathway structure. Z_(M)(x,ω) can beexpressed as:

${Z_{M}( {x,\omega} )} = {\lbrack \frac{R_{e} + R_{i} + R_{g}}{\gamma} \rbrack {\tanh ( {\gamma \; x} )}}$

This equation clearly shows that electrical impedance of the targetpathway structure, and contributions from extracellular fluid resistance(“R_(e)”), intracellular fluid resistance (“R_(i)”) and intermembraneresistance (“R_(g)”) which are electrically connected to target pathwaystructures, all contribute to noise filtering.

A typical approach to evaluation of SNR uses a single value of a rootmean square (RMS) noise voltage. This is calculated by taking a squareroot of an integration of

S_(n)(ω)=4 kT Re[Z_(M)(x,ω)] over all frequencies relevant to eithercomplete membrane response, or to bandwidth of a target pathwaystructure. SNR can be expressed by a ratio:

${SNR} = \frac{{V_{M}(\omega)}}{RMS}$

where |V_(M)(ω)| is maximum amplitude of voltage at each frequency asdelivered by a chosen waveform to the target pathway structure.

An embodiment according to the present invention comprises a pulse burstenvelope having a high spectral density, so that the effect of therapyupon the relevant dielectric pathways, such as, cellular membranereceptors, ion binding to cellular enzymes and general transmembranepotential changes, is enhanced. Accordingly by increasing a number offrequency components transmitted to relevant cellular pathways, a largerange of biophysical phenomena, such as modulating growth factor andcytokine release and ion binding at regulatory molecules, applicable toknown tissue growth mechanisms is accessible. According to an embodimentof the present invention applying a random, or other high spectraldensity envelope, to a pulse burst envelope of mono- or bi-polarrectangular or sinusoidal pulses inducing peak electric fields betweenabout 10⁻⁸ and about 100 V/cm, produces a greater effect on biologicalhealing processes applicable to both soft and hard tissues.

According to yet another embodiment of the present invention by applyinga high spectral density voltage envelope as a modulating or pulse-burstdefining parameter, power requirements for such amplitude modulatedpulse bursts can be significantly lower than that of an unmodulatedpulse burst containing pulses within a similar frequency range. This isdue to a substantial reduction in duty cycle within repetitive bursttrains brought about by imposition of an irregular, and preferablyrandom, amplitude onto what would otherwise be a substantially uniformpulse burst envelope. Accordingly, the dual advantages, of enhancedtransmitted dosimetry to the relevant dielectric pathways and ofdecreased power requirement are achieved.

Referring to FIG. 41, wherein FIG. 41 is a flow diagram of a method fordelivering electromagnetic signals to tissue target pathway structuressuch as ions and ligands of animals, and humans for therapeutic andprophylactic purposes according to an embodiment of the presentinvention. A mathematical model having at least one waveform parameteris applied to configure at least one waveform to be coupled to targetpathway structures such as ions and ligands (Step 41101). The configuredwaveform satisfies a Power SNR model so that for a given and knowntarget pathway structure it is possible to choose at least one waveformparameter so that a waveform is detectable in the target pathwaystructure above its background activity (Step 41102) such as baselinethermal fluctuations in voltage and electrical impedance at a targetpathway structure that depend upon a state of a cell and tissue, that iswhether the state is at least one of resting, growing, replacing, andresponding to injury.

A preferred embodiment of a generated electromagnetic signal iscomprised of a burst of arbitrary waveforms having at least one waveformparameter that includes a plurality of frequency components ranging fromabout 0.01 Hz to about 100 MHz wherein the plurality of frequencycomponents satisfies a Power SNR model (Step 41102). A repetitiveelectromagnetic signal can be generated for example inductively orcapacitively, from said configured at least one waveform (Step 41103).The electromagnetic signal is coupled to a target pathway structure suchas ions and ligands by output of a coupling device such as an electrodeor an inductor, placed in close proximity to the target pathwaystructure (Step 41104) using a positioning device by integrating thecoupling device with the positioning device (Step 41105). The couplingenhances modulation of binding of ions and ligands to regulatorymolecules tissues, cells, and organs. The coupling device can beintegrated into the structure of the positioning device. The positioningdevice can be surgical dressings, wound dressings, pads, seat cushions,mattress pads, shoes, wheelchairs, chairs, and any other garment andstructure that can be juxtaposed to living tissue and cells. Anadvantage of integrating the coupling device with a positioning deviceis that therapeutic treatment can be administered in an unnoticeablefashion and can be administered anywhere and at anytime.

FIG. 42 illustrates a preferred embodiment of an apparatus according tothe present invention. The apparatus is self-contained, lightweight, andportable. A miniature control circuit 42201 is coupled to an end of atleast one connector 42202 such as wire however the control circuit canalso operate wirelessly. The opposite end of the at least one connectoris coupled to a generating device such as an electrical coil 42203. Theminiature control circuit 42201 is constructed in a manner that appliesa mathematical model that is used to configure waveforms. The configuredwaveforms have to satisfy a Power SNR model so that for a given andknown target pathway structure, it is possible to choose waveformparameters that satisfy Power SNR so that a waveform is detectable inthe target pathway structure above its background activity. A preferredembodiment according to the present invention applies a mathematicalmodel to induce a time-varying magnetic field and a time-varyingelectric field in a target pathway structure such as ions and ligands,comprising about 0.1 to about 100 msec bursts of about 1 to about 100microsecond rectangular pulses repeating at about 0.1 to about 100pulses per second. Peak amplitude of the induced electric field isbetween about 1 uV/cm and about 100 mV/cm, varied according to amodified 1/f function where f=frequency. A waveform configured using apreferred embodiment according to the present invention may be appliedto a target pathway structure such as ions and ligands for a preferredtotal exposure time of under 1 minute to 240 minutes daily. Howeverother exposure times can be used. Waveforms configured by the miniaturecontrol circuit 42201 are directed to a generating device 42203 such aselectrical coils via connector 42202. The generating device 42203delivers a pulsing magnetic field configured according to a mathematicalmodel that can be used to provide treatment to a target pathwaystructure such as skin tissue. The miniature control circuit applies apulsing magnetic field for a prescribed time and can automaticallyrepeat applying the pulsing magnetic field for as many applications asare needed in a given time period, for example 10 times a day. Theminiature control circuit can be configured to be programmable applyingpulsing magnetic fields for any time repetition sequence. A preferredembodiment according to the present invention can be positioned to treathair 42204 by being incorporated with a positioning device therebymaking the unit self-contained. Coupling a pulsing magnetic field to atarget pathway structure such as ions and ligands, therapeutically andprophylactically reduces inflammation thereby reducing pain and promoteshealing in treatment areas. When electrical coils are used as thegenerating device 42203, the electrical coils can be powered with a timevarying magnetic field that induces a time varying electric field in atarget pathway structure according to Faraday's law. An electromagneticsignal generated by the generating device 42203 can also be appliedusing electrochemical coupling, wherein electrodes are in direct contactwith skin or another outer electrically conductive boundary of a targetpathway structure. Yet in another embodiment according to the presentinvention, the electromagnetic signal generated by the generating device42203 can also be applied using electrostatic coupling wherein an airgap exists between a generating device 42203 such as an electrode and atarget pathway structure such as ions and ligands. An advantage of thepreferred embodiment according to the present invention is that itsultra lightweight coils and miniaturized circuitry allow for use withcommon physical therapy treatment modalities and at any location forwhich tissue growth, pain relief, and tissue and organ healing isdesired. An advantageous result of application of the preferredembodiment according to the present invention is that tissue growth,repair, and maintenance can be accomplished and enhanced anywhere and atanytime. Yet another advantageous result of application of the preferredembodiment is that growth, repair, and maintenance of molecules, cells,tissues, and organs can be accomplished and enhanced anywhere and atanytime.

FIG. 43 depicts a block diagram of a preferred embodiment according tothe present invention of a miniature control circuit 43300. Theminiature control circuit 43300 produces waveforms that drive agenerating device such as wire coils described above in FIG. 42. Theminiature control circuit can be activated by any activation means suchas an on/off switch. The miniature control circuit 43300 has a powersource such as a lithium battery 43301. A preferred embodiment of thepower source has an output voltage of 3.3 V but other voltages can beused. In another embodiment according to the present invention the powersource can be an external power source such as an electric currentoutlet such as an AC/DC outlet, coupled to the present invention forexample by a plug and wire. A switching power supply 43302 controlsvoltage to a micro-controller 43303. A preferred embodiment of themicro-controller 43303 uses an 8 bit 4 MHz micro-controller 43303 butother bit MHz combination micro-controllers may be used. The switchingpower supply 43302 also delivers current to storage capacitors 43304. Apreferred embodiment of the present invention uses storage capacitorshaving a 220 uF output but other outputs can be used. The storagecapacitors 43304 allow high frequency pulses to be delivered to acoupling device such as inductors (Not Shown). The micro-controller43303 also controls a pulse shaper 43305 and a pulse phase timingcontrol 43306. The pulse shaper 43305 and pulse phase timing control43306 determine pulse shape, burst width, burst envelope shape, andburst repetition rate. An integral waveform generator, such as a sinewave or arbitrary number generator can also be incorporated to providespecific waveforms. A voltage level conversion sub-circuit 43308controls an induced field delivered to a target pathway structure. Aswitching Hexfet 43308 allows pulses of randomized amplitude to bedelivered to output 43309 that routes a waveform to at least onecoupling device such as an inductor. The micro-controller 43303 can alsocontrol total exposure time of a single treatment of a target pathwaystructure such as a molecule, cell, tissue, and organ. The miniaturecontrol circuit 43300 can be constructed to be programmable and apply apulsing magnetic field for a prescribed time and to automatically repeatapplying the pulsing magnetic field for as many applications as areneeded in a given time period, for example 10 times a day. A preferredembodiment according to the present invention uses treatments times ofabout 10 minutes to about 30 minutes. The miniature control circuit43300 can also be integrated with a positioning device. The positioningdevice can also include at least one of a therapeutic surface, atherapeutic structure, and a therapeutic device, such as diathermy,ultrasound, TENS, massage, heat compress, cold compress, anatomicalsupport surfaces, structures, and devices.

Referring to FIG. 44 an embodiment according to the present invention ofa waveform 44400 is illustrated. A pulse 44401 is repeated within aburst 44402 that has a finite duration 44403. The duration 44403 is suchthat a duty cycle which can be defined as a ratio of burst duration tosignal period is between about 1 to about 10⁻⁵. A preferred embodimentaccording to the present invention utilizes pseudo rectangular 10microsecond pulses for pulse 44401 applied in a burst 44402 for about 10to about 50 msec having a modified 1/f amplitude envelope 44404 and witha finite duration 44403 corresponding to a burst period of between about0.1 and about 10 seconds.

Example 1

The Power SNR approach for PMF signal configuration has been testedexperimentally on calcium dependent myosin phosphorylation in a standardenzyme assay. The cell-free reaction mixture was chosen forphosphorylation rate to be linear in time for several minutes, and forsub-saturation Ca²⁺ concentration. This opens the biological window forCa²⁺/CaM to be EMF-sensitive. This system is not responsive to PMF atlevels utilized in this study if Ca²⁺ is at saturation levels withrespect to CaM, and reaction is not slowed to a minute time range.Experiments were performed using myosin light chain (“MLC”) and myosinlight chain kinase (“MLCK”) isolated from turkey gizzard. A reactionmixture consisted of a basic solution containing 40 mM Hepes buffer, pH7.0; 0.5 mM magnesium acetate; 1 mg/ml bovine serum albumin, 0.1% (w/v)Tween 80; and 1 mM EGTA12. Free Ca²⁺ was varied in the 1-7 μM range.Once Ca²⁺ buffering was established, freshly prepared 70 nM CaM, 160 nMMLC and 2 nM MLCK were added to the basic solution to form a finalreaction mixture. The low MLC/MLCK ratio allowed linear time behavior inthe minute time range. This provided reproducible enzyme activities andminimized pipetting time errors.

The reaction mixture was freshly prepared daily for each series ofexperiments and was aliquoted in 100 μL portions into 1.5 ml Eppendorftubes. All Eppendorf tubes containing reaction mixture were kept at 0°C. then transferred to a specially designed water bath maintained at37±0.1° C. by constant perfusion of water prewarmed by passage through aFisher Scientific model 900 heat exchanger. Temperature was monitoredwith a thermistor probe such as a Cole-Parmer model 8110-20, immersed inone Eppendorf tube during all experiments. Reaction was initiated with2.5 μM 32P ATP, and was stopped with Laemmli Sample Buffer solutioncontaining 30 μM EDTA. A minimum of five blank samples were counted ineach experiment. Blanks comprised a total assay mixture minus one of theactive components Ca²⁺, CaM, MLC or MLCK. Experiments for which blankcounts were higher than 300 cpm were rejected. Phosphorylation wasallowed to proceed for 5 min and was evaluated by counting 32Pincorporated in MLC using a TM Analytic model 5303 Mark V liquidscintillation counter.

The signal comprised repetitive bursts of a high frequency waveform.Amplitude was maintained constant at 0.2 G and repetition rate was 1burst/sec for all exposures. Burst duration varied from 65 μsec to 1000μsec based upon projections of Power SNR analysis which showed thatoptimal Power SNR would be achieved as burst duration approached 500μsec. The results are shown in FIG. 7 wherein burst width 701 in μsec isplotted on the x-axis and Myosin Phosphorylation 702 as treated/sham isplotted on the y-axis. It can be seen that the PMF effect on Ca²⁺binding to CaM approaches its maximum at approximately 500 μsec, just asillustrated by the Power SNR model.

These results confirm that a PMF signal, configured according to anembodiment of the present invention, would maximally increase myosinphosphorylation for burst durations sufficient to achieve optimal PowerSNR for a given magnetic field amplitude.

Example 2

According to an embodiment of the present invention use of a Power SNRmodel was further verified in an in vivo wound repair model. A rat woundmodel has been well characterized both biomechanically andbiochemically, and was used in this study. Healthy, young adult maleSprague Dawley rats weighing more than 300 grams were utilized.

The animals were anesthetized with an intraperitoneal dose of Ketamine75 mg/kg and Medetomidine 0.5 mg/kg. After adequate anesthesia had beenachieved, the dorsum was shaved, prepped with a dilute betadine/alcoholsolution, and draped using sterile technique. Using a #10 scalpel, an8-cm linear incision was performed through the skin down to the fasciaon the dorsum of each rat. The wound edges were bluntly dissected tobreak any remaining dermal fibers, leaving an open wound approximately 4cm in diameter. Hemostasis was obtained with applied pressure to avoidany damage to the skin edges. The skin edges were then closed with a 4-0Ethilon running suture. Post-operatively, the animals receivedBuprenorphine 0.1-0.5 mg/kg, intraperitoneal. They were placed inindividual cages and received food and water ad libitum.

PMF exposure comprised two pulsed radio frequency waveforms. The firstwas a standard clinical PRF signal comprising a 65 μsec burst of 27.12MHz sinusoidal waves at 1 Gauss amplitude and repeating at 600bursts/sec. The second was a PRF signal reconfigured according to anembodiment of the present invention. For this signal burst duration wasincreased to 2000 μsec and the amplitude and repetition rate werereduced to 0.2 G and 5 bursts/sec respectively. PRF was applied for 30minutes twice daily.

Tensile strength was performed immediately after wound excision. Two 1cm width strips of skin were transected perpendicular to the scar fromeach sample and used to measure the tensile strength in kg/mm². Thestrips were excised from the same area in each rat to assure consistencyof measurement. The strips were then mounted on a tensiometer. Thestrips were loaded at 10 mm/min and the maximum force generated beforethe wound pulled apart was recorded. The final tensile strength forcomparison was determined by taking the average of the maximum load inkilograms per mm² of the two strips from the same wound.

The results showed average tensile strength for the 65 μsec 1 Gauss PRFsignal was 19.3±4.3 kg/mm² for the exposed group versus 13.0±3.5 kg/mm²for the control group (p<0.01), which is a 48% increase. In contrast,the average tensile strength for the 2000 μsec 0.2 Gauss PRF signal,configured according to an embodiment of the present invention using aPower SNR model was 21.2±5.6 kg/mm² for the treated group versus13.7±4.1 kg/mm² (p<0.01) for the control group, which is a 54% increase.The results for the two signals were not significantly different fromeach other.

These results demonstrate that an embodiment of the present inventionallowed a new PRF signal to be configured that could be produced withsignificantly lower power. The PRF signal configured according to anembodiment of the present invention, accelerated wound repair in the ratmodel in a low power manner versus that for a clinical PRF signal whichaccelerated wound repair but required more than two orders of magnitudemore power to produce.

Example 3

This example illustrates the effects of PRF electromagnetic fieldschosen via the Power SNR method on neurons in culture.

Primary cultures were established from embryonic days 15-16 rodentmesencephalon. This area is dissected, dissociated into single cells bymechanical trituration, and cells are plated in either defined medium ormedium with serum. Cells are typically treated after 6 days of culture,when neurons have matured and developed mechanisms that render themvulnerable to biologically relevant toxins. After treatment, conditionedmedia is collected.

Enzyme linked immunosorbent assays (“ELISAs”) for growth factors such asFibroblast Growth Factor beta (“FGFb”) are used to quantify theirrelease into the medium. Dopaminergic neurons are identified with anantibody to tyrosine hydroxylase (“TH”), an enzyme that converts theamino acid tyrosine to L-dopa, the precursor of dopamine, sincedopaminergic neurons are the only cells that produce this enzyme in thissystem. Cells are quantified by counting TH+ cells in perpendicularstrips across the culture dish under 100× magnification.

Serum contains nutrients and growth factors that support neuronalsurvival. Elimination of serum induces neuronal cell death. Culturemedia was changed and cells were exposed to PMF (power level 6, burstwidth 3000 μsec, and frequency 1 Hz). Four groups were utilized. Group 1used No PMF exposure (null group). Group 2 used Pre-treatment (PMFtreatment 2 hours before medium change). Group 3 used Post-treatment(PMF treatment 2 hours after medium change). Group 4 used Immediatetreatment (PMF treatment simultaneous to medium change).

Results demonstrate a 46% increase in the numbers of survivingdopaminergic neurons after 2 days when cultures were exposed to PMFprior to serum withdrawal. Other treatment regimes had no significanteffects on numbers of surviving neurons. The results are shown in FIG. 6where type of treatment is shown on the x-axis and number of neurons isshown on the y-axis.

FIG. 7, where treatment is shown on the x-axis and number of neurons isshown on the y-axis, illustrates that PMF signals D and E increasenumbers of dopaminergic neurons after reducing serum concentrations inthe medium by 46% and 48% respectively. Both signals were configuredwith a burst width of 3000 μsec, and the repetition rates are 5/sec and1/sec, respectively. Notably, signal D was administered in a chronicparadigm in this experiment, but signal E was administered only once: 2hours prior to serum withdrawal, identical to experiment 1 (see above),producing effects of the same magnitude (46% vs. 48%). Since thereduction of serum in the medium reduces the availability of nutrientsand growth factors, PMF induces the synthesis or release of thesefactors by the cultures themselves.

This portion of the experiment was performed to illustrate the effectsof PMF toxicity induced by 6-OHDA, producing a well-characterizedmechanism of dopaminergic cell death. This molecule enters cells viahigh affinity dopamine transporters and inhibits mitochondrial enzymecomplex I, thus killing these neurons by oxidative stress. Cultures weretreated with 25 μM 6-OHDA after chronic, or acute PMF exposureparadigms. FIG. 8 illustrates these results, where treatment is shown onthe x-axis and number of neurons is shown on the y-axis. The toxinkilled approximately 80% of the dopaminergic neurons in the absence ofPMF treatment. One dose of PMF (power=6; burst width=3000 μsec;frequency=1/sec) significantly increased neuronal survival over 6-OHDAalone (2.6-fold; p≦0.02). This result has particular relevance todeveloping neuroprotection strategies for Parkinson's disease, because6-OHDA is used to lesion dopaminergic neurons in the standard rodentmodel of Parkinson's disease, and the mechanism of toxicity is similarin some ways to the mechanism of neurodegeneration in Parkinson'sdisease itself.

Example 4

In this example electromagnetic field energy was used to stimulateneovascularization in an in vivo model. Two different signal wereemployed, one configured according to prior art and a second configuredaccording to an embodiment of the present invention.

One hundred and eight Sprague-Dawley male rats weighing approximately300 grams each, were equally divided into nine groups. All animals wereanesthetized with a mixture of ketamine/acepromazine/Stadol at 0.1 cc/g.Using sterile surgical techniques, each animal had a 12 cm to 14 cmsegment of tail artery harvested using microsurgical technique. Theartery was flushed with 60 U/ml of heparinized saline to remove anyblood or emboli.

These tail vessels, with an average diameter of 0.4 mm to 0.5 mm, werethen sutured to the transected proximal and distal segments of the rightfemoral artery using two end-to-end anastomoses, creating a femoralarterial loop. The resulting loop was then placed in a subcutaneouspocket created over the animal's abdominal wall/groin musculature, andthe groin incision was closed with 4-0 Ethilon. Each animal was thenrandomly placed into one of nine groups: groups 1 to 3 (controls), theserats received no electromagnetic field treatments and were killed at 4,8, and 12 weeks; groups 4 to 6, 30 min. treatments twice a day using 0.1gauss electromagnetic fields for 4, 8, and 12 weeks (animals were killedat 4, 8, and 12 weeks, respectively); and groups 7 to 9, 30 min.treatments twice a day using 2.0 gauss electromagnetic fields for 4, 8,and 12 weeks (animals were killed at 4, 8, and 12 weeks, respectively).

Pulsed electromagnetic energy was applied to the treated groups using adevice constructed according to an embodiment of the present invention.Animals in the experimental groups were treated for 30 minutes twice aday at either 0.1 gauss or 2.0 gauss, using short pulses (2 msec to 20msec) 27.12 MHz. Animals were positioned on top of the applicator headand confined to ensure that treatment was properly applied. The ratswere reanesthetized with ketamine/acepromazine/Stadol intraperitoneallyand 100 U/kg of heparin intravenously. Using the previous groinincision, the femoral artery was identified and checked for patency. Thefemoral/tail artery loop was then isolated proximally and distally fromthe anastomoses sites, and the vessel was clamped off. Animals were thenkilled. The loop was injected with saline followed by 0.5 cc to 1.0 ccof colored latex through a 25-gauge cannula and clamped. The overlyingabdominal skin was carefully resected, and the arterial loop wasexposed. Neovascularization was quantified by measuring the surface areacovered by new blood-vessel formation delineated by the intraluminallatex. All results were analyzed using the SPSS statistical analysispackage.

The most noticeable difference in neovascularization between treatedversus untreated rats occurred at week 4. At that time, no new vesselformation was found among controls, however, each of the treated groupshad similar statistically significant evidence of neovascularization at0 cm2 versus 1.42±0.80 cm2 (p<0.001). These areas appeared as a latexblush segmentally distributed along the sides of the arterial loop. At 8weeks, controls began to demonstrate neovascularization measured at0.7±0.82 cm2. Both treated groups at 8 weeks again had approximatelyequal statistically significant (p<0.001) outcroppings of blood vesselsof 3.57±1.82 cm2 for the 0.1 gauss group and of 3.77±1.82 cm2 for the2.0 gauss group. At 12 weeks, animals in the control group displayed1.75±0.95 cm2 of neovascularization, whereas the 0.1 gauss groupdemonstrated 5.95±3.25 cm2, and the 2.0 gauss group showed 6.20±3.95 cm2of arborizing vessels. Again, both treated groups displayed comparablestatistically significant findings (p<0.001) over controls.

These experimental findings demonstrate that electromagnetic fieldstimulation of an isolated arterial loop according to an embodiment ofthe present invention increases the amount of quantifiableneovascularization in an in vivo rat model. Increased angiogenesis wasdemonstrated in each of the treated groups at each of the sacrificedates. No differences were found between the results of the two gausslevels tested as predicted by the teachings of the present invention.

Having described embodiments for an integrated coil apparatus fortherapeutically treating human and animal cells, tissues, and organswith electromagnetic fields and method for using same, it is noted thatmodifications and variations can be made by persons skilled in the artin light of the above teachings. It is therefore to be understood thatchanges may be made in the particular embodiments of the inventiondisclosed which are within the scope and spirit of the invention asdefined by the appended claims.

Part 7

Induced time-varying currents from PEMF or PRF devices flow in a hairand cerebrofacial target pathway structure such as a molecule, cell,tissue, and organ, and it is these currents that are a stimulus to whichcells and tissues can react in a physiologically meaningful manner. Theelectrical properties of a hair and cerebrofacial target pathwaystructure affect levels and distributions of induced current. Molecules,cells, tissue, and organs are all in an induced current pathway such ascells in a gap junction contact. Ion or ligand interactions at bindingsites on macromolecules that may reside on a membrane surface arevoltage dependent processes, that is electrochemical, that can respondto an induced electromagnetic field (“E”). Induced current arrives atthese sites via a surrounding ionic medium. The presence of cells in acurrent pathway causes an induced current (“J”) to decay more rapidlywith time (“J(t)”). This is due to an added electrical impedance ofcells from membrane capacitance and time constants of binding and othervoltage sensitive membrane processes such as membrane transport.

Equivalent electrical circuit models representing various membrane andcharged interface configurations have been derived. For example, inCalcium (“Ca²⁺”) binding, the change in concentration of bound Ca²⁺ at abinding site due to induced E may be described in a frequency domain byan impedance expression such as:

${Z_{b}(\omega)} = {R_{ion} + \frac{1}{i\mspace{11mu} \omega \; C_{ion}}}$

which has the form of a series resistance-capacitance electricalequivalent circuit. Where ω is angular frequency defined as 2πf, where fis frequency, i=−1^(1/2), Z_(b)(ω) is the binding impedance, and R_(ion)and C_(ion) are equivalent binding resistance and capacitance of an ionbinding pathway. The value of the equivalent binding time constant,τ_(ion)=R_(ion)C_(ion), is related to a ion binding rate constant,k_(b), via τ_(ion)=R_(ion)C_(ion)=1/k_(b). Thus, the characteristic timeconstant of this pathway is determined by ion binding kinetics.

Induced E from a PEMF or PRF signal can cause current to flow into anion binding pathway and affect the number of Ca²⁺ ions bound per unittime. An electrical equivalent of this is a change in voltage across theequivalent binding capacitance C_(ion), which is a direct measure of thechange in electrical charge stored by C_(ion). Electrical charge isdirectly proportional to a surface concentration of Ca²⁺ ions in thebinding site, that is storage of charge is equivalent to storage of ionsor other charged species on cell surfaces and junctions. Electricalimpedance measurements, as well as direct kinetic analyses of bindingrate constants, provide values for time constants necessary forconfiguration of a PMF waveform to match a bandpass of target pathwaystructures. This allows for a required range of frequencies for anygiven induced E waveform for optimal coupling to target impedance, suchas bandpass.

Ion binding to regulatory molecules is a frequent EMF target, forexample Ca²⁺ binding to calmodulin (“CaM”). Use of this pathway is basedupon acceleration of tissue repair, for example bone repair, woundrepair, hair repair, and repair of other cerebrofacial molecules, cells,tissues, and organs that involves modulation of growth factors releasedin various stages of repair. Growth factors such as platelet derivedgrowth factor (“PDGF”), fibroblast growth factor (“FGF”), and epidermalgrowth factor (“EGF”) are all involved at an appropriate stage ofhealing. Angiogenesis and neovascularization are also integral to tissuegrowth and repair and can be modulated by PMF. All of these factors areCa/CaM-dependent.

Utilizing a Ca/CaM pathway a waveform can be configured for whichinduced power is sufficiently above background thermal noise power.Under correct physiological conditions, this waveform can have aphysiologically significant bioeffect.

Application of a Power SNR model to Ca/CaM requires knowledge ofelectrical equivalents of Ca²⁺ binding kinetics at CaM. Within firstorder binding kinetics, changes in concentration of bound Ca²⁺ at CaMbinding sites over time may be characterized in a frequency domain by anequivalent binding time constant, τ_(ion)=R_(ion)C_(ion), where R_(ion)and C_(ion) are equivalent binding resistance and capacitance of the ionbinding pathway. τ_(ion) is related to a ion binding rate constant,k_(b), via τ_(ion)=R_(ion)C_(ion)=1/k_(b). Published values for k_(b)can then be employed in a cell array model to evaluate SNR by comparingvoltage induced by a PRF signal to thermal fluctuations in voltage at aCaM binding site. Employing numerical values for PMF response, such asV_(max)=6.5×10⁻⁷ sec⁻¹, =2.50/1, K_(D)=30 μM, [Ca²⁺CaM]=K_(D)(+[CaM]),yields k_(b)=665 sec⁻¹ (τ_(ion)=1.5 msec). Such a value for τ_(ion) canbe employed in an electrical equivalent circuit for ion binding whilepower SNR analysis can be performed for any waveform structure.

According to an embodiment of the present invention a mathematical modelfor example a mathematical equation and or a series of mathematicalequations can be configured to assimilate that thermal noise is presentin all voltage dependent processes and represents a minimum thresholdrequirement to establish adequate SNR. For example a mathematical modelthat represents a minimum threshold requirement to establish adequateSNR can be configured to include power spectral density of thermal noisesuch that power spectral density, S_(n)(ω), of thermal noise can beexpressed as:

S _(n)(ω)=4kT Re[Z _(M)(x,ω)]

where Z_(M)(x,ω) is electrical impedance of a target pathway structure,x is a dimension of a target pathway structure and Re denotes a realpart of impedance of a target pathway structure. Z_(M)(x,ω) can beexpressed as:

${Z_{M}( {x,\omega} )} = {\lbrack \frac{R_{e} + R_{i} + R_{g}}{\gamma} \rbrack \tanh \; ( {\gamma \; x} )}$

This equation clearly shows that electrical impedance of the targetpathway structure, and contributions from extracellular fluid resistance(“R_(e)”), intracellular fluid resistance (“R_(i)”) and intermembraneresistance (“R_(g)”) which are electrically connected to a hair andother cerebrofacial target pathway structures, all contribute to noisefiltering.

A typical approach to evaluation of SNR uses a single value of a rootmean square (RMS) noise voltage. This is calculated by taking a squareroot of an integration of

S_(n)(ω)=4 kT Re[Z_(M) (x, ω)] over all frequencies relevant to eithercomplete membrane response, or to bandwidth of a target pathwaystructure. SNR can be expressed by a ratio:

${SNR} = \frac{{V_{M}(\omega)}}{RMS}$

where |V_(M)(ω)| is maximum amplitude of voltage at each frequency asdelivered by a chosen waveform to the target pathway structure.

An embodiment according to the present invention comprises a pulse burstenvelope having a high spectral density, so that the effect of therapyupon the relevant dielectric pathways, such as, cellular membranereceptors, ion binding to cellular enzymes and general transmembranepotential changes, is enhanced. Accordingly by increasing a number offrequency components transmitted to relevant cellular pathways, a largerange of biophysical phenomena, such as modulating growth factor andcytokine release and ion binding at regulatory molecules, applicable toknown hair and other cerebrofacial tissue growth mechanisms isaccessible. According to an embodiment of the present invention applyinga random, or other high spectral density envelope, to a pulse burstenvelope of mono- or bi-polar rectangular or sinusoidal pulses inducingpeak electric fields between about 10⁻⁸ and about 100 V/cm, produces agreater effect on biological healing processes applicable to both softand hard tissues.

According to yet another embodiment of the present invention by applyinga high spectral density voltage envelope as a modulating or pulse-burstdefining parameter, power requirements for such amplitude modulatedpulse bursts can be significantly lower than that of an unmodulatedpulse burst containing pulses within a similar frequency range. This isdue to a substantial reduction in duty cycle within repetitive bursttrains brought about by imposition of an irregular, and preferablyrandom, amplitude onto what would otherwise be a substantially uniformpulse burst envelope. Accordingly, the dual advantages, of enhancedtransmitted dosimetry to the relevant dielectric pathways and ofdecreased power requirement are achieved.

Referring to FIG. 48, wherein FIG. 48 is a flow diagram of a method fordelivering electromagnetic signals that can be pulsed, to hair andcerebrofacial tissue target pathway structures such as ions and ligandsof animals, and humans for therapeutic and prophylactic purposesaccording to an embodiment of the present invention.

At least one waveform having at least one waveform parameter isconfigured to be coupled to hair and cerebrofacial target pathwaystructures such as ions and ligands (Step 48101). Hair and cerebrofacialtarget pathway structures are located in a cerebrofacial treatment area.Examples of a cerebrofacial treatment area include but are not limitedto, hair, a brain, sinuses, adenoids, tonsils, eyes, a nose, ears,teeth, and a tongue.

The at least one waveform parameter is selected to maximize at least oneof a signal to noise ratio and a Power Signal to Noise ratio in a hairand cerebrofacial target pathway structure so that a waveform isdetectable in the hair and cerebofacial target pathway structure aboveits background activity (Step 48102) such as baseline thermalfluctuations in voltage and electrical impedance at a target pathwaystructure that depend upon a state of a cell and tissue, that is whetherthe state is at least one of resting, growing, replacing, and respondingto injury to produce physiologically beneficial results. To bedetectable in the hair and cerebrofacial target pathway structure thevalue of said at least one waveform parameter is chosen by using aconstant of said target pathway structure to evaluate at least one of asignal to noise ratio, and a Power signal to noise ratio, to comparevoltage induced by said at least one waveform in said target pathwaystructure to baseline thermal fluctuations in voltage and electricalimpedance in said target pathway structure whereby bioeffectivemodulation occurs in said target pathway structure by said at least onewaveform by maximizing said at least one of signal to noise ratio andPower signal to noise ratio, within a bandpass of said target pathwaystructure.

A preferred embodiment of a generated electromagnetic signal iscomprised of a burst of arbitrary waveforms having at least one waveformparameter that includes a plurality of frequency components ranging fromabout 0.01 Hz to about 100 MHz wherein the plurality of frequencycomponents satisfies a Power SNR model (Step 48102). A repetitiveelectromagnetic signal can be generated for example inductively orcapacitively, from said configured at least one waveform (Step 48103).The electromagnetic signal can also be non-repetitive. Theelectromagnetic signal is coupled to a hair and cerebrofacial targetpathway structure such as ions and ligands by output of a couplingdevice such as an electrode or an inductor, placed in close proximity tothe target pathway structure (Step 48104). The coupling enhancesmodulation of binding of ions and ligands to regulatory molecules inhair and other cerebrofacial molecules, tissues, cells, and organs.

FIG. 49 illustrates a preferred embodiment of an apparatus according tothe present invention. The apparatus is self-contained, lightweight, andportable. A miniature control circuit 49201 is coupled to an end of atleast one connector 49202 such as wire however the control circuit canalso operate wirelessly. The opposite end of the at least one connectoris coupled to a generating device such as an electrical coil 49203. Theminiature control circuit 49201 is constructed in a manner that appliesa mathematical model that is used to configure waveforms. The configuredwaveforms have to satisfy Power SNR so that for a given and known hairand cerebrofacial target pathway structure, it is possible to choosewaveform parameters that satisfy Power SNR so that a waveform producesphysiologically beneficial results, for example bioeffective modulation,and is detectable in the hair and cerebrofacial target pathway structureabove its background activity. A preferred embodiment according to thepresent invention applies a mathematical model to induce a time-varyingmagnetic field and a time-varying electric field in a hair andcerebrofacial target pathway structure such as ions and ligands,comprising about 0.1 to about 100 msec bursts of about 1 to about 100microsecond rectangular pulses repeating at about 0.1 to about 100pulses per second. Peak amplitude of the induced electric field isbetween about 1 uV/cm and about 100 mV/cm, varied according to amodified 1/f function where f=frequency. A waveform configured using apreferred embodiment according to the present invention may be appliedto a hair and cerebrofacial target pathway structure such as ions andligands for a preferred total exposure time of under 1 minute to 240minutes daily. However other exposure times can be used. Waveformsconfigured by the miniature control circuit 49201 are directed to agenerating device 49203 such as electrical coils via connector 49202.The generating device 49203 delivers a pulsing magnetic field that canbe used to provide treatment to a hair and cerebrofacial target pathwaystructure such as hair tissue. The miniature control circuit applies apulsing magnetic field for a prescribed time and can automaticallyrepeat applying the pulsing magnetic field for as many applications asare needed in a given time period, for example 10 times a day. Theminiature control circuit can be configured to be programmable applyingpulsing magnetic fields for any time repetition sequence. A preferredembodiment according to the present invention can be positioned to treathair 49204 by being incorporated into a positioning device therebymaking the unit self-contained. Coupling a pulsing magnetic field to ahair and cerebrofacial target pathway structure such as ions andligands, therapeutically and prophylactically reduces inflammationthereby advantageously reducing pain and promoting healing incerebrofacial areas. When electrical coils are used as the generatingdevice 49203, the electrical coils can be powered with a time varyingmagnetic field that induces a time varying electric field in a targetpathway structure according to Faraday's law. An electromagnetic signalgenerated by the generating device 49203 can also be applied usingelectrochemical coupling, wherein electrodes are in direct contact withskin or another outer electrically conductive boundary of a hair andcerebrofacial target pathway structure. Yet in another embodimentaccording to the present invention, the electromagnetic signal generatedby the generating device 49203 can also be applied using electrostaticcoupling wherein an air gap exists between a generating device 49203such as an electrode and a hair and cerebrofacial target pathwaystructure such as ions and ligands. An advantage of the preferredembodiment according to the present invention is that its ultralightweight coils and miniaturized circuitry allow for use with commonphysical therapy treatment modalities and at any cerebrofacial locationfor which hair growth, pain relief, and tissue and organ healing isdesired. An advantageous result of application of the preferredembodiment according to the present invention is that hair growth,repair, and maintenance can be accomplished and enhanced anywhere and atanytime, for example while driving a car or watching television. Yetanother advantageous result of application of the preferred embodimentis that growth, repair, and maintenance of cerebrofacial molecules,cells, tissues, and organs can be accomplished and enhanced anywhere andat anytime, for example while driving a car or watching television.

FIG. 50 depicts a block diagram of a preferred embodiment according tothe present invention of a miniature control circuit 50300. Theminiature control circuit 50300 produces waveforms that drive agenerating device such as wire coils described above in FIG. 49. Theminiature control circuit can be activated by any activation means suchas an on/off switch. The miniature control circuit 50300 has a powersource such as a lithium battery 50301. A preferred embodiment of thepower source has an output voltage of 3.3 V but other voltages can beused. In another embodiment according to the present invention the powersource can be an external power source such as an electric currentoutlet such as an AC/DC outlet, coupled to the present invention forexample by a plug and wire. A switching power supply 50302 controlsvoltage to a micro-controller 50303. A preferred embodiment of themicro-controller 50303 uses an 8 bit 4 MHz micro-controller 50303 butother bit MHz combination micro-controllers may be used. The switchingpower supply 50302 also delivers current to storage capacitors 50304. Apreferred embodiment of the present invention uses storage capacitorshaving a 220 uF output but other outputs can be used. The storagecapacitors 50304 allow high frequency pulses to be delivered to acoupling device such as inductors (Not Shown). The micro-controller50303 also controls a pulse shaper 50305 and a pulse phase timingcontrol 50306. The pulse shaper 50305 and pulse phase timing control50306 determine pulse shape, burst width, burst envelope shape, andburst repetition rate. An integral waveform generator, such as a sinewave or arbitrary number generator can also be incorporated to providespecific waveforms. A voltage level conversion sub-circuit 50307controls an induced field delivered to a target pathway structure. Aswitching Hexfet 50308 allows pulses of randomized amplitude to bedelivered to output 50309 that routes a waveform to at least onecoupling device such as an inductor. The micro-controller 50303 can alsocontrol total exposure time of a single treatment of a hair andcerebrofacial target pathway structure such as a molecule, cell, tissue,and organ. The miniature control circuit 50300 can be constructed to beprogrammable and apply a pulsing magnetic field for a prescribed timeand to automatically repeat applying the pulsing magnetic field for asmany applications as are needed in a given time period, for example 10times a day. A preferred embodiment according to the present inventionuses treatments times of about 10 minutes to about 30 minutes.

Referring to FIG. 51 an embodiment according to the present invention ofa waveform 51400 is illustrated. A pulse 51401 is repeated within aburst 51402 that has a finite duration 51403. The duration 51403 is suchthat a duty cycle which can be defined as a ratio of burst duration tosignal period is between about 1 to about 10⁻⁵. A preferred embodimentaccording to the present invention utilizes pseudo rectangular 10microsecond pulses for pulse 51401 applied in a burst 51402 for about 10to about 50 msec having a modified 1/f amplitude envelope 51404 and witha finite duration 51403 corresponding to a burst period of between about0.1 and about 10 seconds.

Example 1

The Power SNR approach for PMF signal configuration has been testedexperimentally on calcium dependent myosin phosphorylation in a standardenzyme assay. The cell-free reaction mixture was chosen forphosphorylation rate to be linear in time for several minutes, and forsub-saturation Ca²⁺ concentration. This opens the biological window forCa²⁺/CaM to be EMF-sensitive. This system is not responsive to PMF atlevels utilized in this study if Ca²⁺ is at saturation levels withrespect to CaM, and reaction is not slowed to a minute time range.Experiments were performed using myosin light chain (“MLC”) and myosinlight chain kinase (“MLCK”) isolated from turkey gizzard. A reactionmixture consisted of a basic solution containing 40 mM Hepes buffer, pH7.0; 0.5 mM magnesium acetate; 1 mg/ml bovine serum albumin, 0.1% (w/v)Tween 80; and 1 mM EGTA12. Free Ca²⁺ was varied in the 1-7 μM range.Once Ca²⁺ buffering was established, freshly prepared 70 nM CaM, 160 nMMLC and 2 nM MLCK were added to the basic solution to form a finalreaction mixture. The low MLC/MLCK ratio allowed linear time behavior inthe minute time range. This provided reproducible enzyme activities andminimized pipetting time errors.

The reaction mixture was freshly prepared daily for each series ofexperiments and was aliquoted in 100 μL portions into 1.5 ml Eppendorftubes. All Eppendorf tubes containing reaction mixture were kept at 0°C. then transferred to a specially designed water bath maintained at37±0.1° C. by constant perfusion of water prewarmed by passage through aFisher Scientific model 900 heat exchanger. Temperature was monitoredwith a thermistor probe such as a Cole-Parmer model 8110-20, immersed inone Eppendorf tube during all experiments. Reaction was initiated with2.5 μM 32P ATP, and was stopped with Laemmli Sample Buffer solutioncontaining 30 μM EDTA. A minimum of five blank samples were counted ineach experiment. Blanks comprised a total assay mixture minus one of theactive components Ca²⁺, CaM, MLC or MLCK. Experiments for which blankcounts were higher than 300 cpm were rejected. Phosphorylation wasallowed to proceed for 5 min and was evaluated by counting 32Pincorporated in MLC using a TM Analytic model 5303 Mark V liquidscintillation counter.

The signal comprised repetitive bursts of a high frequency waveform.Amplitude was maintained constant at 0.2 G and repetition rate was 1burst/sec for all exposures. Burst duration varied from 65 μsec to 1000μsec based upon projections of Power SNR analysis which showed thatoptimal Power SNR would be achieved as burst duration approached 500μsec. The results are shown in FIG. 52 wherein burst width 52501 in μsecis plotted on the x-axis and Myosin Phosphorylation 52502 astreated/sham is plotted on the y-axis. It can be seen that the PMFeffect on Ca²⁺ binding to CaM approaches its maximum at approximately500 μsec, just as illustrated by the Power SNR model.

These results confirm that a PMF signal, configured according to anembodiment of the present invention, would maximally increase myosinphosphorylation for burst durations sufficient to achieve optimal PowerSNR for a given magnetic field amplitude.

Example 2

According to an embodiment of the present invention use of a Power SNRmodel was further verified in an in vivo wound repair model. A rat woundmodel has been well characterized both biomechanically andbiochemically, and was used in this study. Healthy, young adult maleSprague Dawley rats weighing more than 300 grams were utilized.

The animals were anesthetized with an intraperitoneal dose of Ketamine75 mg/kg and Medetomidine 0.5 mg/kg. After adequate anesthesia had beenachieved, the dorsum was shaved, prepped with a dilute betadine/alcoholsolution, and draped using sterile technique. Using a #10 scalpel, an8-cm linear incision was performed through the skin down to the fasciaon the dorsum of each rat. The wound edges were bluntly dissected tobreak any remaining dermal fibers, leaving an open wound approximately 4cm in diameter. Hemostasis was obtained with applied pressure to avoidany damage to the skin edges. The skin edges were then closed with a 4-0Ethilon running suture. Post-operatively, the animals receivedBuprenorphine 0.1-0.5 mg/kg, intraperitoneal. They were placed inindividual cages and received food and water ad libitum.

PMF exposure comprised two pulsed radio frequency waveforms. The firstwas a standard clinical PRF signal comprising a 65 μsec burst of 27.12MHz sinusoidal waves at 1 Gauss amplitude and repeating at 600bursts/sec. The second was a PRF signal reconfigured according to anembodiment of the present invention. For this signal burst duration wasincreased to 2000 μsec and the amplitude and repetition rate werereduced to 0.2 G and 5 bursts/sec respectively. PRF was applied for 30minutes twice daily.

Tensile strength was performed immediately after wound excision. Two 1cm width strips of skin were transected perpendicular to the scar fromeach sample and used to measure the tensile strength in kg/mm². Thestrips were excised from the same area in each rat to assure consistencyof measurement. The strips were then mounted on a tensiometer. Thestrips were loaded at 10 mm/min and the maximum force generated beforethe wound pulled apart was recorded. The final tensile strength forcomparison was determined by taking the average of the maximum load inkilograms per mm² of the two strips from the same wound.

The results showed average tensile strength for the 65 μsec 1 Gauss PRFsignal was 19.3±4.3 kg/mm² for the exposed group versus 13.0±3.5 kg/mm²for the control group (p<0.01), which is a 48% increase. In contrast,the average tensile strength for the 2000 μsec 0.2 Gauss PRF signal,configured according to an embodiment of the present invention using aPower SNR model was 21.2±5.6 kg/mm² for the treated group versus13.7±4.1 kg/mm² (p<0.01) for the control group, which is a 54% increase.The results for the two signals were not significantly different fromeach other.

These results demonstrate that an embodiment of the present inventionallowed a new PRF signal to be configured that could be produced withsignificantly lower power. The PRF signal configured according to anembodiment of the present invention, accelerated wound repair in the ratmodel in a low power manner versus that for a clinical PRF signal whichaccelerated wound repair but required more than two orders of magnitudemore power to produce.

Example 3

This example illustrates the effects of PRF electromagnetic fieldschosen via the Power SNR method on neurons in culture.

Primary cultures were established from embryonic days 15-16 rodentmesencephalon. This area is dissected, dissociated into single cells bymechanical trituration, and cells are plated in either defined medium ormedium with serum. Cells are typically treated after 6 days of culture,when neurons have matured and developed mechanisms that render themvulnerable to biologically relevant toxins. After treatment, conditionedmedia is collected.

Enzyme linked immunosorbent assays (“ELISAs”) for growth factors such asFibroblast Growth Factor beta (“FGFb”) are used to quantify theirrelease into the medium. Dopaminergic neurons are identified with anantibody to tyrosine hydroxylase (“TH”), an enzyme that converts theamino acid tyrosine to L-dopa, the precursor of dopamine, sincedopaminergic neurons are the only cells that produce this enzyme in thissystem. Cells are quantified by counting TH+ cells in perpendicularstrips across the culture dish under 100× magnification.

Serum contains nutrients and growth factors that support neuronalsurvival. Elimination of serum induces neuronal cell death. Culturemedia was changed and cells were exposed to PMF (power level 6, burstwidth 3000 μsec, and frequency 1 Hz). Four groups were utilized. Group 1used No PMF exposure (null group). Group 2 used Pre-treatment (PMFtreatment 2 hours before medium change). Group 3 used Post-treatment(PMF treatment 2 hours after medium change). Group 4 used Immediatetreatment (PMF treatment simultaneous to medium change).

Results demonstrate a 46% increase in the numbers of survivingdopaminergic neurons after 2 days when cultures were exposed to PMFprior to serum withdrawal. Other treatment regimes had no significanteffects on numbers of surviving neurons. The results are shown in FIG.53 where type of treatment is shown on the x-axis and number of neuronsis shown on the y-axis.

FIG. 53, where treatment 53601 is shown on the x-axis and number ofneurons 53602 is shown on the y-axis, illustrates that PMF signals D andE increase numbers of dopaminergic neurons after reducing serumconcentrations in the medium by 46% and 48% respectively. Both signalswere configured with a burst width of 3000 μsec, and the repetitionrates are 5/sec and 1/sec, respectively. Notably, signal D wasadministered in a chronic paradigm in this experiment, but signal E wasadministered only once: 2 hours prior to serum withdrawal, identical toexperiment 1 (see above), producing effects of the same magnitude (46%vs. 48%). Since the reduction of serum in the medium reduces theavailability of nutrients and growth factors, PMF induces the synthesisor release of these factors by the cultures themselves.

This portion of the experiment was performed to illustrate the effectsof PMF toxicity induced by 6-OHDA, producing a well-characterizedmechanism of dopaminergic cell death. This molecule enters cells viahigh affinity dopamine transporters and inhibits mitochondrial enzymecomplex I, thus killing these neurons by oxidative stress. Cultures weretreated with 25 μM 6-hydroxydopamine (“6-OHDA”) after chronic, or acutePMF exposure paradigms. FIG. 54 illustrates these results, wheretreatment 54701 is shown on the x-axis and number of neurons 54702 isshown on the y-axis. The toxin killed approximately 80% of thedopaminergic neurons in the absence of PMF treatment. One dose of PMF(power=6; burst width=3000 μsec; frequency=1/sec) significantlyincreased neuronal survival over 6-OHDA alone (2.6-fold; p<0.02). Thisresult has particular relevance to developing neuroprotection strategiesfor Parkinson's disease, because 6-OHDA is used to lesion dopaminergicneurons in the standard rodent model of Parkinson's disease, and themechanism of toxicity is similar in some ways to the mechanism ofneurodegeneration in Parkinson's disease itself.

Example 4

In this example electromagnetic field energy was used to stimulateneovascularization in an in vivo model. Two different signal wereemployed, one configured according to prior art and a second configuredaccording to an embodiment of the present invention.

One hundred and eight Sprague-Dawley male rats weighing approximately300 grams each, were equally divided into nine groups. All animals wereanesthetized with a mixture of ketamine/acepromazine/Stadol at 0.1 cc/g.Using sterile surgical techniques, each animal had a 12 cm to 14 cmsegment of tail artery harvested using microsurgical technique. Theartery was flushed with 60 U/ml of heparinized saline to remove anyblood or emboli.

These tail vessels, with an average diameter of 0.4 mm to 0.5 mm, werethen sutured to the transected proximal and distal segments of the rightfemoral artery using two end-to-end anastomoses, creating a femoralarterial loop. The resulting loop was then placed in a subcutaneouspocket created over the animal's abdominal wall/groin musculature, andthe groin incision was closed with 4-0 Ethilon. Each animal was thenrandomly placed into one of nine groups: groups 1 to 3 (controls), theserats received no electromagnetic field treatments and were killed at 4,8, and 12 weeks; groups 4 to 6, 30 min. treatments twice a day using 0.1gauss electromagnetic fields for 4, 8, and 12 weeks (animals were killedat 4, 8, and 12 weeks, respectively); and groups 7 to 9, 30 min.treatments twice a day using 2.0 gauss electromagnetic fields for 4, 8,and 12 weeks (animals were killed at 4, 8, and 12 weeks, respectively).

Pulsed electromagnetic energy was applied to the treated groups using adevice constructed according to an embodiment of the present invention.Animals in the experimental groups were treated for 30 minutes twice aday at either 0.1 gauss or 2.0 gauss, using short pulses (2 msec to 20msec) 27.12 MHz. Animals were positioned on top of the applicator headand confined to ensure that treatment was properly applied. The ratswere reanesthetized with ketamine/acepromazine/Stadol intraperitoneallyand 100 U/kg of heparin intravenously. Using the previous groinincision, the femoral artery was identified and checked for patency. Thefemoral/tail artery loop was then isolated proximally and distally fromthe anastomoses sites, and the vessel was clamped off. Animals were thenkilled. The loop was injected with saline followed by 0.5 cc to 1.0 ccof colored latex through a 25-gauge cannula and clamped. The overlyingabdominal skin was carefully resected, and the arterial loop wasexposed. Neovascularization was quantified by measuring the surface areacovered by new blood-vessel formation delineated by the intraluminallatex. All results were analyzed using the SPSS statistical analysispackage.

The most noticeable difference in neovascularization between treatedversus untreated rats occurred at week 4. At that time, no new vesselformation was found among controls, however, each of the treated groupshad similar statistically significant evidence of neovascularization at0 cm2 versus 1.42±0.80 cm2 (p<0.001). These areas appeared as a latexblush segmentally distributed along the sides of the arterial loop. At 8weeks, controls began to demonstrate neovascularization measured at0.7±0.82 cm2. Both treated groups at 8 weeks again had approximatelyequal statistically significant (p<0.001) outcroppings of blood vesselsof 3.57±1.82 cm2 for the 0.1 gauss group and of 3.77±1.82 cm2 for the2.0 gauss group. At 12 weeks, animals in the control group displayed1.75±0.95 cm2 of neovascularization, whereas the 0.1 gauss groupdemonstrated 5.95±3.25 cm2, and the 2.0 gauss group showed 6.20±3.95 cm2of arborizing vessels. Again, both treated groups displayed comparablestatistically significant findings (p<0.001) over controls.

These experimental findings demonstrate that electromagnetic fieldstimulation of an isolated arterial loop according to an embodiment ofthe present invention increases the amount of quantifiableneovascularization in an in vivo rat model. Increased angiogenesis wasdemonstrated in each of the treated groups at each of the sacrificedates. No differences were found between the results of the two gausslevels tested as predicted by the teachings of the present invention.

Having described embodiments for an apparatus and a method for treatmentof hair restoration and cerebrofacial conditions that is self-containedand delivers electromagnetic treatment to hair and other cerebrofacialtissue, it is noted that modifications and variations can be made bypersons skilled in the art in light of the above teachings. It istherefore to be understood that changes may be made in the particularembodiments of the invention disclosed which are within the scope andspirit of the invention as defined by the appended claims.

Part 8

Induced time-varying currents from PEMF or PRF devices flow in a targetpathway structure such as a molecule, cell, tissue, and organ, and it isthese currents that are a stimulus to which cells and tissues can reactin a physiologically meaningful manner. The electrical properties of atarget pathway structure affect levels and distributions of inducedcurrent. Molecules, cells, tissue, and organs are all in an inducedcurrent pathway such as cells in a gap junction contact. Ion or ligandinteractions at binding sites on macromolecules that may reside on amembrane surface are voltage dependent processes, that iselectrochemical, that can respond to an induced electromagnetic field(“E”). Induced current arrives at these sites via a surrounding ionicmedium. The presence of cells in a current pathway causes an inducedcurrent (“J”) to decay more rapidly with time (“J(t)”). This is due toan added electrical impedance of cells from membrane capacitance and ionbinding time constants of binding and other voltage sensitive membraneprocesses such as membrane transport. Knowledge of ion binding timeconstants allows SNR to be evaluated for any EMF signal configuration. Apreferred embodiment according to the present invention uses ion bindingtime constants in the range of about 1 to about 100 msec.

Equivalent electrical circuit models representing various membrane andcharged interface configurations have been derived. For example, inCalcium (“Ca2+”) binding, the change in concentration of bound Ca²⁺ at abinding site due to induced E may be described in a frequency domain byan impedance expression such as:

${Z_{b}(\omega)} = {R_{ion} + \frac{1}{i\; \omega \; C_{ion}}}$

which has the form of a series resistance-capacitance electricalequivalent circuit. Where ω is angular frequency defined as 2πf, where fis frequency, i=−1^(1/2), Z_(b)(ω) is the binding impedance, and R_(ion)and C_(ion) are equivalent binding resistance and capacitance of an ionbinding pathway. The value of the equivalent binding time constant,τ_(ion)=R_(ion)C_(ion), is related to a ion binding rate constant,k_(b), via τ_(ion)=R_(ion)C_(ion)=1/k_(b). Thus, the characteristic timeconstant of this pathway is determined by ion binding kinetics.

Induced E from a PEMF or PRF signal can cause current to flow into anion binding pathway and affect the number of Ca²⁺ ions bound per unittime. An electrical equivalent of this is a change in voltage across theequivalent binding capacitance C_(ion), which is a direct measure of thechange in electrical charge stored by C_(ion). Electrical charge isdirectly proportional to a surface concentration of Ca²⁺ ions in thebinding site, that is storage of charge is equivalent to storage of ionsor other charged species on cell surfaces and junctions. Electricalimpedance measurements, as well as direct kinetic analyses of bindingrate constants, provide values for time constants necessary forconfiguration of a PMF waveform to match a bandpass of target pathwaystructures. This allows for a required range of frequencies for anygiven induced E waveform for optimal coupling to target impedance, suchas bandpass.

Ion binding to regulatory molecules is a frequent EMF target, forexample Ca²⁺ binding to calmodulin (“CaM”). Use of this pathway is basedupon acceleration of tissue repair, for example bone repair, woundrepair, hair repair, and repair of other molecules, cells, tissues, andorgans that involves modulation of growth factors released in variousstages of repair. Growth factors such as platelet derived growth factor(“PDGF”), fibroblast growth factor (“FGF”), and epidermal growth factor(“EGF”) are all involved at an appropriate stage of healing.Angiogenesis and neovascularization are also integral to tissue growthand repair and can be modulated by PMF. All of these factors areCa/CaM-dependent.

Utilizing a Ca/CaM pathway a waveform can be configured for whichinduced power is sufficiently above background thermal noise power.Under correct physiological conditions, this waveform can have aphysiologically significant bioeffect.

Application of a Power SNR model to Ca/CaM requires knowledge ofelectrical equivalents of Ca²⁺ binding kinetics at CaM. Within firstorder binding kinetics, changes in concentration of bound Ca²⁺ at CaMbinding sites over time may be characterized in a frequency domain by anequivalent binding time constant, τ_(ion)=R_(ion)C_(ion), where R_(ion)and C_(ion) are equivalent binding resistance and capacitance of the ionbinding pathway. τ_(ion) is related to a ion binding rate constant,k_(b), via τ_(ion)=R_(ion)C_(ion)=1/k_(b). Published values for k_(b)can then be employed in a cell array model to evaluate SNR by comparingvoltage induced by a PRF signal to thermal fluctuations in voltage at aCaM binding site. Employing numerical values for PMF response, such asV_(max)=6.5×10⁻⁷ sec⁻¹, =2.50/1, K_(D)=30 μM, [Ca²⁺CaM]=K_(D)(+[CaM]),yields k_(b)=665 sec⁻¹ (τ_(ion)=1.5 msec). Such a value for τ_(ion) canbe employed in an electrical equivalent circuit for ion binding whilepower SNR analysis can be performed for any waveform structure.

According to an embodiment of the present invention a mathematical modelcan be configured to assimilate that thermal noise is present in allvoltage dependent processes and represents a minimum thresholdrequirement to establish adequate SNR. Power spectral density, S_(n)(ω),of thermal noise can be expressed as:

S _(n)(ω)=4kT Re[Z _(M)(x,ω)]

where Z_(M)(x, ω) is electrical impedance of a target pathway structure,x is a dimension of a target pathway structure and Re denotes a realpart of impedance of a target pathway structure. Z_(M)(x, ω) can beexpressed as:

${Z_{M}( {x,\omega} )} = {\lbrack \frac{R_{e} + R_{i} + R_{g}}{y} \rbrack \tanh \; ( {y\; x} )}$

This equation clearly shows that electrical impedance of the targetpathway structure, and contributions from extracellular fluid resistance(“R_(e)”), intracellular fluid resistance (“R_(i)”) and intermembraneresistance (“R_(g)”) which are electrically connected to a targetpathway structure, all contribute to noise filtering.

A typical approach to evaluation of SNR uses a single value of a rootmean square (RMS) noise voltage. This is calculated by taking a squareroot of an integration of S_(n)(ω)=4 kT Re[Z_(M)(x, ω)] over allfrequencies relevant to either complete membrane response, or tobandwidth of a target pathway structure. SNR can be expressed by aratio:

${SNR} = \frac{{V_{M}(\omega)}}{RMS}$

where |V_(M)(ω)| is maximum amplitude of voltage at each frequency asdelivered by a chosen waveform to the target pathway structure.

An embodiment according to the present invention comprises a pulse burstenvelope having a high spectral density, so that the effect of therapyupon the relevant dielectric pathways, such as, cellular membranereceptors, ion binding to cellular enzymes and general transmembranepotential changes, is enhanced. Accordingly by increasing a number offrequency components transmitted to relevant cellular pathways, a largerange of biophysical phenomena, such as modulating growth factor andcytokine release and ion binding at regulatory molecules, applicable toknown tissue growth mechanisms is accessible. According to an embodimentof the present invention applying a random, or other high spectraldensity envelope, to a pulse burst envelope of mono-polar or bi-polarrectangular or sinusoidal pulses inducing peak electric fields betweenabout 10⁻⁸ and about 100 V/cm, produces a greater effect on biologicalhealing processes applicable to both soft and hard tissues.

According to yet another embodiment of the present invention by applyinga high spectral density voltage envelope as a modulating or pulse-burstdefining parameter, power requirements for such amplitude modulatedpulse bursts can be significantly lower than that of an unmodulatedpulse burst containing pulses within a similar frequency range. This isdue to a substantial reduction in duty cycle within repetitive bursttrains brought about by imposition of an irregular amplitude andpreferably a random amplitude onto what would otherwise be asubstantially uniform pulse burst envelope. Accordingly, the dualadvantages, of enhanced transmitted dosimetry to the relevant dielectricpathways and of decreased power requirement are achieved.

Referring to FIG. 55 wherein FIG. 55 is a flow diagram of a method forgenerating electromagnetic signals to be coupled to an eye according toan embodiment of the present invention, a target pathway structure suchas ions and ligands, is identified. Establishing a baseline backgroundactivity such as baseline thermal fluctuations in voltage and electricalimpedance, at the target pathway structure by determining a state of atleast one of a cell and a tissue at the target pathway structure,wherein the state is at least one of resting, growing, replacing, andresponding to injury. (STEP 55101) The state of the at least one of acell and a tissue is determined by its response to injury or insult.Configuring at least one waveform to have sufficient signal to noiseratio to modulate at least one of ion and ligand interactions wherebythe at least one of ion and ligand interactions are detectable in thetarget pathway structure above the established baseline thermalfluctuations in voltage and electrical impedance. (STEP 55102)Generating an electromagnetic signal from the configured at least onewaveform. (STEP 55103) The electromagnetic signal can be generated byusing at least one waveform configured by applying a mathematical modelsuch as an equation, formula, or function having at least one waveformparameter that satisfies an SNR or Power SNR mathematical model suchthat ion and ligand interactions are modulated and the at least oneconfigured waveform is detectable at the target pathway structure aboveits established background activity. Coupling the electromagnetic signalto the target pathway structure using a coupling device. (STEP 55104)The generated electromagnetic signals can be coupled for therapeutic andprophylactic purposes. Since ophthalmic tissue is very delicate,application of electromagnetic signals using an embodiment according tothe present invention is extremely safe and efficient since theapplication of electromagnetic signals is non-invasive.

A preferred embodiment of a generated electromagnetic signal iscomprised of a burst of arbitrary waveforms having at least one waveformparameter that includes a plurality of frequency components ranging fromabout 0.01 Hz to about 100 MHz wherein the plurality of frequencycomponents satisfies a Power SNR model. A repetitive electromagneticsignal can be generated for example inductively or capacitively, fromthe configured at least one waveform. The electromagnetic signal iscoupled to a target pathway structure such as ions and ligands by outputof a coupling device such as an electrode or an inductor, placed inclose proximity to the target pathway structure using a positioningdevice. The coupling enhances modulation of binding of ions and ligandsto regulatory molecules tissues, cells, and organs. According to anembodiment of the present invention EMF signals configured using SNRanalysis to match the bandpass of a second messenger whereby the EMFsignals can act as a first messenger to modulate biochemical cascadessuch as production of cytokines, Nitric Oxide, Nitric Oxide Synthase andgrowth factors that are related to tissue growth and repair. Adetectable E field amplitude is produced within a frequency response ofCa²⁺ binding.

FIG. 56 illustrates a preferred embodiment of an apparatus according tothe present invention. The apparatus is self-contained, lightweight, andportable. A miniature control circuit 56201 is coupled to an end of atleast one connector 56202 such as wire however the control circuit canalso operate wirelessly. The opposite end of the at least one connectoris coupled to a generating device such as an electrical coil 56203. Theminiature control circuit 56201 is constructed in a manner that appliesa mathematical model that is used to configure waveforms. The configuredwaveforms have to satisfy a Power SNR model so that for a given andknown target pathway structure, it is possible to choose waveformparameters that satisfy Power SNR so that a waveform is detectable inthe target pathway structure above its background activity. A preferredembodiment according to the present invention applies a mathematicalmodel to induce a time-varying magnetic field and a time-varyingelectric field in a target pathway structure such as ions and ligands,comprising about 0.001 to about 100 msec bursts of about 1 to about 100microsecond rectangular pulses repeating at about 0.1 to about 100pulses per second. Peak amplitude of the induced electric field isbetween about 1 uV/cm and about 100 mV/cm, varied according to amodified 1/f function where f=frequency. A waveform configured using apreferred embodiment according to the present invention may be appliedto a target pathway structure such as ions and ligands for a preferredtotal exposure time of under 1 minute to 240 minutes daily. Howeverother exposure times can be used. Waveforms configured by the miniaturecontrol circuit 56201 are directed to a generating device 56203 such aselectrical coils via connector 56202. The generating device 56203delivers a pulsing magnetic field configured according to a mathematicalmodel that can be used to provide treatment to a target pathwaystructure such as eye tissue. The miniature control circuit applies apulsing magnetic field for a prescribed time and can automaticallyrepeat applying the pulsing magnetic field for as many applications asare needed in a given time period, for example 10 times a day. Theminiature control circuit can be configured to be programmable applyingpulsing magnetic fields for any time repetition sequence. A preferredembodiment according to the present invention can be positioned to treatophthalmic tissue by being incorporated with a positioning device 56204such as an eye-patch, eyeglasses, goggles, and monocles thereby makingthe unit self-contained. Coupling a pulsing magnetic field to a targetpathway structure such as ions and ligands, therapeutically andprophylactically reduces inflammation thereby reducing pain and promoteshealing in treatment areas. When electrical coils are used as thegenerating device 56203, the electrical coils can be powered with a timevarying magnetic field that induces a time varying electric field in atarget pathway structure according to Faraday's law. An electromagneticsignal generated by the generating device 203 can also be applied usingelectrochemical coupling, wherein electrodes are in direct contact withskin or another outer electrically conductive boundary of a targetpathway structure. Yet in another embodiment according to the presentinvention, the electromagnetic signal generated by the generating device56203 can also be applied using electrostatic coupling wherein an airgap exists between a generating device 56203 such as an electrode and atarget pathway structure such as ions and ligands. An advantage of thepreferred embodiment according to the present invention is that itsultra lightweight coils and miniaturized circuitry allow for use withcommon physical therapy treatment modalities and at any location forwhich tissue growth, pain relief, and tissue and organ healing isdesired. An advantageous result of application of the preferredembodiment according to the present invention is that tissue growth,repair, and maintenance can be accomplished and enhanced anywhere and atanytime. Yet another advantageous result of application of the preferredembodiment is that growth, repair, and maintenance of molecules, cells,tissues, and organs can be accomplished and enhanced anywhere and atanytime. A preferred embodiment according to the present inventiondelivers PEMF for application to ophthalmic tissue that is infected withdiseases as macular degeneration, glaucoma, retinosa pigmentosa, repairand regeneration of optic nerve prophylaxis, and other related diseases.

FIG. 57 depicts a block diagram of a preferred embodiment according tothe present invention of a miniature control circuit 57300. Theminiature control circuit 57300 produces waveforms that drive agenerating device such as wire coils described above in FIG. 56. Theminiature control circuit can be activated by any activation means suchas an on/off switch. The miniature control circuit 57300 has a powersource such as a lithium battery 57301. A preferred embodiment of thepower source has an output voltage of 3.3 V but other voltages can beused. In another embodiment according to the present invention the powersource can be an external power source such as an electric currentoutlet such as an AC/DC outlet, coupled to the present invention forexample by a plug and wire. A switching power supply 57302 controlsvoltage to a micro-controller 57303. A preferred embodiment of themicro-controller 57303 uses an 8 bit 4 MHz micro-controller 57303 butother bit MHz combination micro-controllers may be used. The switchingpower supply 57302 also delivers current to storage capacitors 57304. Apreferred embodiment of the present invention uses storage capacitorshaving a 220 uF output but other outputs can be used. The storagecapacitors 57304 allow high frequency pulses to be delivered to acoupling device such as inductors (Not Shown). The micro-controller57303 also controls a pulse shaper 57305 and a pulse phase timingcontrol 306. The pulse shaper 57305 and pulse phase timing control 57306determine pulse shape, burst width, burst envelope shape, and burstrepetition rate. In a preferred embodiment according to the presentinvention the pulse shaper 57305 and phase timing control 57306 areconfigured such that the waveforms configured are detectable abovebackground activity at a target pathway structure by satisfying at leastone of a SNR and Power SNR mathematical model. An integral waveformgenerator, such as a sine wave or arbitrary number generator can also beincorporated to provide specific waveforms. A voltage level conversionsub-circuit 57308 controls an induced field delivered to a targetpathway structure. A switching Hexfet 57308 allows pulses of randomizedamplitude to be delivered to output 57309 that routes a waveform to atleast one coupling device such as an inductor. The micro-controller57303 can also control total exposure time of a single treatment of atarget pathway structure such as a molecule, cell, tissue, and organ.The miniature control circuit 57300 can be constructed to beprogrammable and apply a pulsing magnetic field for a prescribed timeand to automatically repeat applying the pulsing magnetic field for asmany applications as are needed in a given time period, for example 10times a day. A preferred embodiment according to the present inventionuses treatments times of about 10 minutes to about 30 minutes.

FIG. 58 depicts a block diagram of an embodiment according to thepresent invention of a miniature control circuit 58400. The miniaturecontrol circuit 58400 produces waveforms that drive a generating devicesuch as wire coils described above in FIG. 56. The miniature controlcircuit can be activated by any activation means such as an on/offswitch. The miniature control circuit 58400 has a power source such as alithium battery 58401. In another embodiment according to the presentinvention the power source can be an external power source such as anelectric current outlet such as an AC/DC outlet, coupled to the presentinvention for example by a plug and wire. A user input/output means58402 such as an on/off switch controls voltage to the miniature controlcircuit and is connected to a cpu-control 58403. The cpu-control 58403creates a SNR EMF waveform by processing information provided to it viaflash memory programmed having SNR EMF signal parameters such as pulseshape, burst width, burst envelope shape, and burst repetition rate. Thewaveform is pulse modulated by a modulator 58405 interfacing with anoscillator 58406 having a crystal 58407 controlled by the cpu-control58403 according to the SNR EMF signal parameters programmed into theflash memory of the cpu-control 58403. The oscillator 58406 having acrystal 58407 provides a carrier frequency. A preferred embodiment ofthe crystal is a 27.120 MHz crystal but other MHz crystals can be used.The modulated waveform is then amplified by an amp 58408 and sent to anoutput stage means 58409 where the amplified modulated waveform ismatched to impedance via a R C circuit across a patient applicator 58410such as a coil. The patient applicator generates a SNR EMF signal to bedelivered to a patient.

Referring to FIG. 59 an embodiment according to the present invention ofa waveform 59500 is illustrated. A pulse 59501 is repeated within aburst 59502 that has a finite duration 59503. The duration 59503 is suchthat a duty cycle which can be defined as a ratio of burst duration tosignal period is between about 1 to about 10⁻⁵. A preferred embodimentaccording to the present invention utilizes pseudo rectangular 10microsecond pulses for pulse 59501 applied in a burst 59502 for about 10to about 50 msec having a modified 1/f amplitude envelope 59504 and witha finite duration 59503 corresponding to a burst period of between about0.1 and about 10 seconds.

It is further intended that any other embodiments of the presentinvention that result from any changes in application or method of useor operation, method of manufacture, shape, size or material which arenot specified within the detailed written description or illustrationsand drawings contained herein, yet are considered apparent or obvious toone skilled in the art, are within the scope of the present invention.

Example 1

The Power SNR approach for PMF signal configuration has been testedexperimentally on calcium dependent myosin phosphorylation in a standardenzyme assay. The cell-free reaction mixture was chosen forphosphorylation rate to be linear in time for several minutes, and forsub-saturation Ca²⁺ concentration. This opens the biological window forCa²⁺/CaM to be EMF-sensitive. This system is not responsive to PMF atlevels utilized in this study if Ca²⁺ is at saturation levels withrespect to CaM, and reaction is not slowed to a minute time range.Experiments were performed using myosin light chain (“MLC”) and myosinlight chain kinase (“MLCK”) isolated from turkey gizzard. A reactionmixture consisted of a basic solution containing 40 mM Hepes buffer, pH7.0; 0.5 mM magnesium acetate; 1 mg/ml bovine serum albumin, 0.1% (w/v)Tween 80; and 1 mM EGTA12. Free Ca²⁺ was varied in the 1-7 μM range.Once Ca²⁺ buffering was established, freshly prepared 70 nM CaM, 160 nMMLC and 2 nM MLCK were added to the basic solution to form a finalreaction mixture. The low MLC/MLCK ratio allowed linear time behavior inthe minute time range. This provided reproducible enzyme activities andminimized pipetting time errors.

The reaction mixture was freshly prepared daily for each series ofexperiments and was aliquoted in 100 μL portions into 1.5 ml Eppendorftubes. All Eppendorf tubes containing reaction mixture were kept at 0°C. then transferred to a specially designed water bath maintained at37±0.1° C. by constant perfusion of water prewarmed by passage through aFisher Scientific model 900 heat exchanger. Temperature was monitoredwith a thermistor probe such as a Cole-Parmer model 8110-20, immersed inone Eppendorf tube during all experiments. Reaction was initiated with2.5 μM 32P ATP, and was stopped with Laemmli Sample Buffer solutioncontaining 30 μM EDTA. A minimum of five blank samples were counted ineach experiment. Blanks comprised a total assay mixture minus one of theactive components Ca²⁺, CaM, MLC or MLCK. Experiments for which blankcounts were higher than 300 cpm were rejected. Phosphorylation wasallowed to proceed for 5 min and was evaluated by counting 32Pincorporated in MLC using a TM Analytic model 5303 Mark V liquidscintillation counter.

The signal comprised repetitive bursts of a high frequency waveform.Amplitude was maintained constant at 0.2 G and repetition rate was 1burst/sec for all exposures. Burst duration varied from 65 μsec to 1000μsec based upon projections of Power SNR analysis which showed thatoptimal Power SNR would be achieved as burst duration approached 500μsec. The results are shown in FIG. 60 wherein burst width 60601 in μsecis plotted on the x-axis and Myosin Phosphorylation 60602 astreated/sham is plotted on the y-axis. It can be seen that the PMFeffect on Ca²⁺ binding to CaM approaches its maximum at approximately500 μsec, just as illustrated by the Power SNR model.

These results confirm that a PMF signal, configured according to anembodiment of the present invention, would maximally increase myosinphosphorylation for burst durations sufficient to achieve optimal PowerSNR for a given magnetic field amplitude.

Example 2

According to an embodiment of the present invention use of a Power SNRmodel was further verified in an in vivo wound repair model. A rat woundmodel has been well characterized both biomechanically andbiochemically, and was used in this study. Healthy, young adult maleSprague Dawley rats weighing more than 300 grams were utilized.

The animals were anesthetized with an intraperitoneal dose of Ketamine75 mg/kg and Medetomidine 0.5 mg/kg. After adequate anesthesia had beenachieved, the dorsum was shaved, prepped with a dilute betadine/alcoholsolution, and draped using sterile technique. Using a #10 scalpel, an8-cm linear incision was performed through the skin down to the fasciaon the dorsum of each rat. The wound edges were bluntly dissected tobreak any remaining dermal fibers, leaving an open wound approximately 4cm in diameter. Hemostasis was obtained with applied pressure to avoidany damage to the skin edges. The skin edges were then closed with a 4-0Ethilon running suture. Post-operatively, the animals receivedBuprenorphine 0.1-0.5 mg/kg, intraperitoneal. They were placed inindividual cages and received food and water ad libitum.

PMF exposure comprised two pulsed radio frequency waveforms. The firstwas a standard clinical PRF signal comprising a 65 μsec burst of 27.12MHz sinusoidal waves at 1 Gauss amplitude and repeating at 600bursts/sec. The second was a PRF signal reconfigured according to anembodiment of the present invention. For this signal burst duration wasincreased to 2000 μsec and the amplitude and repetition rate werereduced to 0.2 G and 5 bursts/sec respectively. PRF was applied for 30minutes twice daily.

Tensile strength was performed immediately after wound excision. Two 1cm width strips of skin were transected perpendicular to the scar fromeach sample and used to measure the tensile strength in kg/mm2. Thestrips were excised from the same area in each rat to assure consistencyof measurement. The strips were then mounted on a tensiometer. Thestrips were loaded at 10 mm/min and the maximum force generated beforethe wound pulled apart was recorded. The final tensile strength forcomparison was determined by taking the average of the maximum load inkilograms per mm2 of the two strips from the same wound.

The results showed average tensile strength for the 65 μsec 1 Gauss PRFsignal was 19.3±4.3 kg/mm2 for the exposed group versus 13.0±3.5 kg/mm2for the control group (p<0.01), which is a 48% increase. In contrast,the average tensile strength for the 2000 μsec 0.2 Gauss PRF signal,configured according to an embodiment of the present invention using aPower SNR model was 21.2±5.6 kg/mm2 for the treated group versus13.7±4.1 kg/mm2 (p<0.01) for the control group, which is a 54% increase.The results for the two signals were not significantly different fromeach other.

These results demonstrate that an embodiment of the present inventionallowed a new PRF signal to be configured that could be produced withsignificantly lower power. The PRF signal configured according to anembodiment of the present invention, accelerated wound repair in the ratmodel in a low power manner versus that for a clinical PRF signal whichaccelerated wound repair but required more than two orders of magnitudemore power to produce.

Example 3

This example illustrates the effects of PRF electromagnetic fieldschosen via the Power SNR method on neurons in culture.

Primary cultures were established from embryonic days 15-16 rodentmesencephalon. This area is dissected, dissociated into single cells bymechanical trituration, and cells are plated in either defined medium ormedium with serum. Cells are typically treated after 6 days of culture,when neurons have matured and developed mechanisms that render themvulnerable to biologically relevant toxins. After treatment, conditionedmedia is collected.

Enzyme linked immunosorbent assays (“ELISAs”) for growth factors such asFibroblast Growth Factor beta (“FGFb”) are used to quantify theirrelease into the medium. Dopaminergic neurons are identified with anantibody to tyrosine hydroxylase (“TH”), an enzyme that converts theamino acid tyrosine to L-dopa, the precursor of dopamine, sincedopaminergic neurons are the only cells that produce this enzyme in thissystem. Cells are quantified by counting TH+ cells in perpendicularstrips across the culture dish under 100.times. magnification.

Serum contains nutrients and growth factors that support neuronalsurvival. Elimination of serum induces neuronal cell death. Culturemedia was changed and cells were exposed to PMF (power level 6, burstwidth 3000 μsec, and frequency 1 Hz). Four groups were utilized. Group 1used No PMF exposure (null group). Group 2 used Pre-treatment (PMFtreatment 2 hours before medium change). Group 3 used Post-treatment(PMF treatment 2 hours after medium change). Group 4 used Immediatetreatment (PMF treatment simultaneous to medium change).

Results demonstrate a 46% increase in the numbers of survivingdopaminergic neurons after 2 days when cultures were exposed to PMFprior to serum withdrawal. Other treatment regimes had no significanteffects on numbers of surviving neurons. The results are shown in FIG.60 where type of treatment 60601 is shown on the x-axis and number ofneurons 60602 is shown on the y-axis.

FIG. 61, where treatment 61701 is shown on the x-axis and number ofneurons 61702 is shown on the y-axis, illustrates that PMF signals D andE increase numbers of dopaminergic neurons after reducing serumconcentrations in the medium by 46% and 48% respectively. Both signalswere configured with a burst width of 3000 μsec, and the repetitionrates are 5/sec and 1/sec, respectively. Notably, signal D wasadministered in a chronic paradigm in this experiment, but signal E wasadministered only once: 2 hours prior to serum withdrawal, identical toexperiment 1 (see above), producing effects of the same magnitude (46%vs. 48%). Since the reduction of serum in the medium reduces theavailability of nutrients and growth factors, PMF induces the synthesisor release of these factors by the cultures themselves.

This portion of the experiment was performed to illustrate the effectsof PMF toxicity induced by 6-OHDA, producing a well-characterizedmechanism of dopaminergic cell death. This molecule enters cells viahigh affinity dopamine transporters and inhibits mitochondrial enzymecomplex I, thus killing these neurons by oxidative stress. Cultures weretreated with 25 μM 6-OHDA after chronic, or acute PMF exposureparadigms. FIG. 62 illustrates these results, where treatment 62801 isshown on the x-axis and number of neurons 62802 is shown on the y-axis.The toxin killed approximately 80% of the dopaminergic neurons in theabsence of PMF treatment. One dose of PMF (power=6; burst width=3000μsec; frequency=1/sec) significantly increased neuronal survival over6-OHDA alone (2.6-fold; p≦0.02). This result has particular relevance todeveloping neuroprotection strategies for Parkinson's disease, because6-OHDA is used to lesion dopaminergic neurons in the standard rodentmodel of Parkinson's disease, and the mechanism of toxicity is similarin some ways to the mechanism of neurodegeneration in Parkinson'sdisease itself.

Example 4

In this example electromagnetic field energy was used to stimulateneovascularization in an in vivo model. Two different signal wereemployed, one configured according to prior art and a second configuredaccording to an embodiment of the present invention.

One hundred and eight Sprague-Dawley male rats weighing approximately300 grams each, were equally divided into nine groups. All animals wereanesthetized with a mixture of ketamine/acepromazine/Stadol at 0.1 cc/g.Using sterile surgical techniques, each animal had a 12 cm to 14 cmsegment of tail artery harvested using microsurgical technique. Theartery was flushed with 60 U/ml of heparinized saline to remove anyblood or emboli.

These tail vessels, with an average diameter of 0.4 mm to 0.5 mm, werethen sutured to the transected proximal and distal segments of the rightfemoral artery using two end-to-end anastomoses, creating a femoralarterial loop. The resulting loop was then placed in a subcutaneouspocket created over the animal's abdominal wall/groin musculature, andthe groin incision was closed with 4-0 Ethilon. Each animal was thenrandomly placed into one of nine groups: groups 1 to 3 (controls), theserats received no electromagnetic field treatments and were killed at 4,8, and 12 weeks; groups 4 to 6, 30 min. treatments twice a day using 0.1gauss electromagnetic fields for 4, 8, and 12 weeks (animals were killedat 4, 8, and 12 weeks, respectively); and groups 7 to 9, 30 min.treatments twice a day using 2.0 gauss electromagnetic fields for 4, 8,and 12 weeks (animals were killed at 4, 8, and 12 weeks, respectively).

Pulsed electromagnetic energy was applied to the treated groups using adevice constructed according to an embodiment of the present invention.Animals in the experimental groups were treated for 30 minutes twice aday at either 0.1 gauss or 2.0 gauss, using short pulses (2 msec to 20msec) 27.12 MHz. Animals were positioned on top of the applicator headand confined to ensure that treatment was properly applied. The ratswere reanesthetized with ketamine/acepromazine/Stadol intraperitoneallyand 100 U/kg of heparin intravenously. Using the previous groinincision, the femoral artery was identified and checked for patency. Thefemoral/tail artery loop was then isolated proximally and distally fromthe anastomoses sites, and the vessel was clamped off. Animals were thenkilled. The loop was injected with saline followed by 0.5 cc to 1.0 ccof colored latex through a 25-gauge cannula and clamped. The overlyingabdominal skin was carefully resected, and the arterial loop wasexposed. Neovascularization was quantified by measuring the surface areacovered by new blood-vessel formation delineated by the intraluminallatex. All results were analyzed using the SPSS statistical analysispackage.

The most noticeable difference in neovascularization between treatedversus untreated rats occurred at week 4. At that time, no new vesselformation was found among controls, however, each of the treated groupshad similar statistically significant evidence of neovascularization at0 cm2 versus 1.42±0.80 cm2 (p<0.001). These areas appeared as a latexblush segmentally distributed along the sides of the arterial loop. At 8weeks, controls began to demonstrate neovascularization measured at0.7±0.82 cm2. Both treated groups at 8 weeks again had approximatelyequal statistically significant (p<0.001) outcroppings of blood vesselsof 3.57±1.82 cm2 for the 0.1 gauss group and of 3.77±1.82 cm2 for the2.0 gauss group. At 12 weeks, animals in the control group displayed1.75±0.95 cm2 of neovascularization, whereas the 0.1 gauss groupdemonstrated 5.95±3.25 cm2, and the 2.0 gauss group showed 6.20±3.95 cm2of arborizing vessels. Again, both treated groups displayed comparablestatistically significant findings (p<0.001) over controls.

These experimental findings demonstrate that electromagnetic fieldstimulation of an isolated arterial loop according to an embodiment ofthe present invention increases the amount of quantifiableneovascularization in an in vivo rat model. Increased angiogenesis wasdemonstrated in each of the treated groups at each of the sacrificedates. No differences were found between the results of the two gausslevels tested as predicted by the teachings of the present invention.

Having described embodiments for an apparatus for applyingelectromagnetic signals to an eye and method for using same, it is notedthat modifications and variations can be made by persons skilled in theart in light of the above teachings. It is therefore to be understoodthat changes may be made in the particular embodiments of the inventiondisclosed which are within the scope and spirit of the invention asdefined by the appended claims.

Part 9

Induced time-varying currents from PEMF or PRF devices flow in a targetpathway structure such as a molecule, cell, tissue, and organ, and it isthese currents that are a stimulus to which cells and tissues can reactin a physiologically meaningful manner. The electrical properties of atarget pathway structure affect levels and distributions of inducedcurrent. Molecules, cells, tissue, and organs are all in an inducedcurrent pathway such as cells in a gap junction contact. Ion or ligandinteractions at binding sites on macromolecules that may reside on amembrane surface are voltage dependent processes, that iselectrochemical, that can respond to an induced electromagnetic field(“E”). Induced current arrives at these sites via a surrounding ionicmedium. The presence of cells in a current pathway causes an inducedcurrent (“J”) to decay more rapidly with time (“J(t)”). This is due toan added electrical impedance of cells from membrane capacitance and ionbinding time constants of binding and other voltage sensitive membraneprocesses such as membrane transport. Knowledge of ion binding timeconstants allows SNR to be evaluated for any EMF signal configuration.Preferably ion binding time constants in the range of about 1 to about100 msec are used.

Equivalent electrical circuit models representing various membrane andcharged interface configurations have been derived. For example, inCalcium (“Ca²⁺”) binding, the change in concentration of bound Ca²⁺ at abinding site due to induced E may be described in a frequency domain byan impedance expression such as:

${Z_{b}\mspace{11mu} (\omega)} = {R_{ion} + \frac{1}{i\mspace{11mu} \omega \; C_{ion}}}$

which has the form of a series resistance-capacitance electricalequivalent circuit. Where ω is angular frequency defined as 2πf, where fis frequency, i=−1^(1/2), Z_(b)(ω) is the binding impedance, and R_(ion)and C_(ion) are equivalent binding resistance and capacitance of an ionbinding pathway. The value of the equivalent binding time constant,τ_(ion)=R_(ion)C_(ion), is related to a ion binding rate constant,k_(b), via τ_(ion)=R_(ion)C_(ion)=1/k_(b). Thus, the characteristic timeconstant of this pathway is determined by ion binding kinetics.

Induced E from a PEMF or PRF signal can cause current to flow into anion binding pathway and affect the number of Ca²⁺ ions bound per unittime. An electrical equivalent of this is a change in voltage across theequivalent binding capacitance C_(ion), which is a direct measure of thechange in electrical charge stored by Cion. Electrical charge isdirectly proportional to a surface concentration of Ca²⁺ ions in thebinding site that is storage of charge is equivalent to storage of ionsor other charged species on cell surfaces and junctions. Electricalimpedance measurements, as well as direct kinetic analyses of bindingrate constants, provide values for time constants necessary forconfiguration of a PMF waveform to match a bandpass of target pathwaystructures. This allows for a required range of frequencies for anygiven induced E waveform for optimal coupling to target impedance, suchas bandpass.

Ion binding to regulatory molecules is a frequent EMF target, forexample Ca²⁺ binding to calmodulin (“CaM”). Use of this pathway is basedupon acceleration of tissue repair, for example bone repair, woundrepair, hair repair, and repair of other molecules, cells, tissues, andorgans that involves modulation of growth factors released in variousstages of repair. Growth factors such as platelet derived growth factor(“PDGF”), fibroblast growth factor (“FGF”), and epidermal growth factor(“EGF”) are all involved at an appropriate stage of healing.Angiogenesis and neovascularization are also integral to tissue growthand repair and can be modulated by PMF. All of these factors areCa/CaM-dependent.

Utilizing a Ca/CaM pathway a waveform can be configured for whichinduced power is sufficiently above background thermal noise power.Under correct physiological conditions, this waveform can have aphysiologically significant bioeffect.

Application of a Power SNR model to Ca/CaM requires knowledge ofelectrical equivalents of Ca2+ binding kinetics at CaM. Within firstorder binding kinetics, changes in concentration of bound Ca²⁺ at CaMbinding sites over time may be characterized in a frequency domain by anequivalent binding time constant, τ_(ion)=R_(ion)C_(ion), where R_(ion)and C_(ion) are equivalent binding resistance and capacitance of the ionbinding pathway. τ_(ion) is related to a ion binding rate constant,k_(b), via τ_(ion)=R_(ion)C_(ion)=1/k_(b). Published values for k_(b)can then be employed in a cell array model to evaluate SNR by comparingvoltage induced by a PRF signal to thermal fluctuations in voltage at aCaM binding site. Employing numerical values for PMF response, such asVmax=6.5×10−7 sec⁻¹, =2.504, K_(D)=30 μM, [Ca²⁺CaM]=K_(D)(+[CaM]),yields k_(b)=665 sec⁻¹ (τ_(ion)=1.5 msec). Such a value for τ_(ion) canbe employed in an electrical equivalent circuit for ion binding whilepower SNR analysis can be performed for any waveform structure.

According to an embodiment of the present invention a mathematical modelcan be configured to assimilate that thermal noise is present in allvoltage dependent processes and represents a minimum thresholdrequirement to establish adequate SNR. Power spectral density, S_(n)(ω),of thermal noise can be expressed as:

S _(n)(ω)=4kT Re[Z _(M)(x,ω)]

where Z_(M)(x,ω) is electrical impedance of a target pathway structure,x is a dimension of a target pathway structure and Re denotes a realpart of impedance of a target pathway structure. Z_(M)(x,ω) can beexpressed as:

${Z_{M}( {x,\omega} )} = {\lbrack \frac{R_{e} + R_{i} + R_{g}}{\gamma} \rbrack \tanh \; ( {\gamma \; x} )}$

This equation clearly shows that electrical impedance of the targetpathway structure, and contributions from extracellular fluid resistance(“Re”), intracellular fluid resistance (“Ri”) and intermembraneresistance (“Rg”) which are electrically connected to target pathwaystructures all contribute to noise filtering.

A typical approach to evaluation of SNR uses a single value of a rootmean square (RMS) noise voltage. This is calculated by taking a squareroot of an integration of S_(n)(ω)=4 kT Re[Z_(M) (x, ω)] over allfrequencies relevant to either a complete membrane response, or tobandwidth of a target pathway structure. SNR can be expressed by aratio:

${SNR} = \frac{{V_{M}(\omega)}}{RMS}$

where |V_(M)(ω)| is maximum amplitude of voltage at each frequency asdelivered by a chosen waveform to the target pathway structure.

An embodiment according to the present invention comprises a pulse burstenvelope having a high spectral density, so that the effect of therapyupon the relevant dielectric pathways, such as, cellular membranereceptors, ion binding to cellular enzymes and general transmembranepotential changes, is enhanced. Accordingly by increasing a number offrequency components transmitted to relevant cellular pathways, a largerange of biophysical phenomena, such as modulating growth factor andcytokine release and ion binding at regulatory molecules, applicable toknown tissue growth mechanisms is accessible. According to an embodimentof the present invention applying a random, or other high spectraldensity envelope, to a pulse burst envelope of mono-polar or bi-polarrectangular or sinusoidal pulses inducing peak electric fields betweenabout 10⁻⁸ and about 100 V/cm, produces a greater effect on biologicalhealing processes applicable to both soft and hard tissues.

According to yet another embodiment of the present invention by applyinga high spectral density voltage envelope as a modulating or pulse-burstdefining parameter, power requirements for such amplitude modulatedpulse bursts can be significantly lower than that of an unmodulatedpulse burst containing pulses within a similar frequency range. This isdue to a substantial reduction in duty cycle within repetitive bursttrains brought about by imposition of an irregular amplitude andpreferably a random amplitude onto what would otherwise be asubstantially uniform pulse burst envelope. Accordingly, the dualadvantages, of enhanced transmitted dosimetry to the relevant dielectricpathways and of decreased power requirement are achieved.

Referring to FIG. 63 wherein FIG. 63 is a flow diagram of a method forgenerating electromagnetic signals to be coupled to a respiratory targetpathway structure according to an embodiment of the present invention, atarget pathway structure such as ions and ligands, is identified.Establishing a baseline background activity such as baseline thermalfluctuations in voltage and electrical impedance, at the target pathwaystructure by determining a state of at least one of a cell and a tissueat the target pathway structure, wherein the state is at least one ofresting, growing, replacing, and responding to injury. (STEP 63101) Thestate of the at least one of a cell and a tissue is determined by itsresponse to injury or insult. Configuring at least one waveform to havesufficient signal to noise ratio to modulate at least one of ion andligand interactions whereby the at least one of ion and ligandinteractions are detectable in the target pathway structure above theestablished baseline thermal fluctuations in voltage and electricalimpedance. (STEP 63102) Repetitively generating an electromagneticsignal from the configured at least one waveform. (STEP 63103) Theelectromagnetic signal can be generated by using at least one waveformconfigured by applying a mathematical model such as an equation,formula, or function having at least one waveform parameter thatsatisfies an SNR or Power SNR mathematical model such that ion andligand interactions are modulated and the at least one configuredwaveform is detectable at the target pathway structure above itsestablished background activity. Coupling the electromagnetic signal tothe target pathway structure using a coupling device. (STEP 63104) Thegenerated electromagnetic signals can be coupled for therapeutic andprophylactic purposes. The coupling enhances a stimulus that cells andtissues react to in a physiological meaningful manner for example,treatment of lung diseases resulting from inflammatory processes causedby inhalation of foreign material into lung tissue. Since lung tissue isvery delicate, application of electromagnetic signals using anembodiment according to the present invention is extremely safe andefficient since the application of electromagnetic signals isnon-invasive.

In an aspect of the present invention, a generated electromagneticsignal is comprised of a burst of arbitrary waveforms having at leastone waveform parameter that includes a plurality of frequency componentsranging from about 0.01 Hz to about 100 MHz wherein the plurality offrequency components satisfies a Power SNR model. A repetitiveelectromagnetic signal can be generated for example inductively orcapacitively, from the configured at least one waveform. Theelectromagnetic signal is coupled to a target pathway structure such asions and ligands by output of a coupling device such as an electrode oran inductor, placed in close proximity to the target pathway structureusing a positioning device. The coupling enhances modulation of bindingof ions and ligands to regulatory molecules, tissues, cells, and organs.According to an embodiment of the present invention EMF signalsconfigured using SNR analysis to match the bandpass of a secondmessenger whereby the EMF signals can act as a first messenger tomodulate biochemical cascades such as production of cytokines, NitricOxide, Nitric Oxide Synthase and growth factors that are related totissue growth and repair. A detectable E field amplitude is producedwithin a frequency response of Ca²⁺ binding.

FIG. 64 illustrates an embodiment of an apparatus according to thepresent invention. The apparatus is self-contained, lightweight, andportable. A miniature control circuit 64201 is connected to a generatingdevice such as an electrical coil 64202. The miniature control circuit64201 is constructed in a manner that applies a mathematical model thatis used to configure waveforms. The configured waveforms have to satisfya Power SNR model so that for a given and known target pathwaystructure, it is possible to choose waveform parameters that satisfyPower SNR so that a waveform is detectable in the target pathwaystructure above its background activity. An embodiment according to thepresent invention applies a mathematical model to induce a time-varyingmagnetic field and a time-varying electric field in a target pathwaystructure such as ions and ligands, comprising about 0.001 to about 100msec bursts of about 1 to about 100 microsecond rectangular pulsesrepeating at about 0.1 to about 100 pulses per second. Peak amplitude ofthe induced electric field is between about 1 uV/cm and about 100 mV/cm,varied according to a modified 1/f function where f=frequency. Awaveform configured using an embodiment according to the presentinvention may be applied to a target pathway structure such as ions andligands, preferably for a total exposure time of under 1 minute to 240minutes daily. However other exposure times can be used. Waveformsconfigured by the miniature control circuit 64201 are directed to agenerating device 64202 such as electrical coils. Preferably, thegenerating device 64202 is a comformable coil for example pliable,comprising one or more turns of electrically conducting wire in agenerally circular or oval shape however other shapes can be used. Thegenerating device 64202 delivers a pulsing magnetic field configuredaccording to a mathematical model that can be used to provide treatmentto a target pathway structure such as lung tissue. The miniature controlcircuit applies a pulsing magnetic field for a prescribed time and canautomatically repeat applying the pulsing magnetic field for as manyapplications as are needed in a given time period, for example 12 timesa day. The miniature control circuit can be configured to beprogrammable applying pulsing magnetic fields for any time repetitionsequence. An embodiment according to the present invention can bepositioned to treat respiratory tissue by being incorporated with apositioning device such as a bandage or a vest thereby making the unitself-contained. Coupling a pulsing magnetic field to a target pathwaystructure such as ions and ligands, therapeutically and prophylacticallyreduces inflammation thereby reducing pain and promotes healing intreatment areas. When electrical coils are used as the generating device64202, the electrical coils can be powered with a time varying magneticfield that induces a time varying electric field in a target pathwaystructure according to Faraday's law. An electromagnetic signalgenerated by the generating device 64202 can also be applied usingelectrochemical coupling, wherein electrodes are in direct contact withskin or another outer electrically conductive boundary of a targetpathway structure. Yet in another embodiment according to the presentinvention, the electromagnetic signal generated by the generating device64202 can also be applied using electrostatic coupling wherein an airgap exists between a generating device 64202 such as an electrode and atarget pathway structure such as ions and ligands. An advantage of thepresent invention is that its ultra lightweight coils and miniaturizedcircuitry allow for use with common physical therapy treatmentmodalities, and at any location for which tissue growth, pain relief,and tissue and organ healing is desired. An advantageous result ofapplication of the present invention is that tissue growth, repair, andmaintenance can be accomplished and enhanced anywhere and at anytime.Yet another advantageous result of application of the present inventionis that growth, repair, and maintenance of molecules, cells, tissues,and organs can be accomplished and enhanced anywhere and at anytime.Another embodiment according to the present invention delivers PEMF forapplication to respiratory tissue that is infected with diseases such assarcoidosis, granulomatous pneumonitis, pulmonary fibrosis, and “WorldTrade Center Cough.”

FIG. 65 depicts a block diagram of an embodiment according to thepresent invention of a miniature control circuit 65300. The miniaturecontrol circuit 65300 produces waveforms that drive a generating devicesuch as wire coils described above in FIG. 64. The miniature controlcircuit can be activated by any activation means such as an on/offswitch. The miniature control circuit 65300 has a power source such as alithium battery 65301. Preferably the power source has an output voltageof 3.3 V but other voltages can be used. In another embodiment accordingto the present invention the power source can be an external powersource such as an electric current outlet such as an AC/DC outlet,coupled to the present invention for example by a plug and wire. Aswitching power supply 65302 controls voltage to a micro-controller 303.Preferably the micro-controller 65303 uses an 8 bit 4 MHzmicro-controller 65303 but other bit MHz combination micro-controllersmay be used. The switching power supply 65302 also delivers current tostorage capacitors 65304. Preferably the storage capacitors 65304 havinga 220 uF output but other outputs can be used. The storage capacitors65304 allow high frequency pulses to be delivered to a coupling devicesuch as inductors (Not Shown). The micro-controller 65303 also controlsa pulse shaper 65305 and a pulse phase timing control 65306. The pulseshaper 65305 and pulse phase timing control 65306 determine pulse shape,burst width, burst envelope shape, and burst repetition rate. In anaspect of the present invention the pulse shaper 65305 and phase timingcontrol 65306 are configured such that the waveforms configured aredetectable above background activity at a target pathway structure bysatisfying at least one of a SNR and Power SNR mathematical model. Anintegral waveform generator, such as a sine wave or arbitrary numbergenerator can also be incorporated to provide specific waveforms. Avoltage level conversion sub-circuit 307 controls an induced fielddelivered to a target pathway structure. A switching Hexfet 65308 allowspulses of randomized amplitude to be delivered to output 65309 thatroutes a waveform to at least one coupling device such as an inductor.The micro-controller 65303 can also control total exposure time of asingle treatment of a target pathway structure such as a molecule, cell,tissue, and organ. The miniature control circuit 65300 can beconstructed to be programmable and apply a pulsing magnetic field for aprescribed time and to automatically repeat applying the pulsingmagnetic field for as many applications as are needed in a given timeperiod, for example 10 times a day. Preferably treatments times of about1 minutes to about 30 minutes are used.

Referring to FIG. 66 an embodiment according to the present invention ofa waveform 66400 is illustrated. A pulse 66401 is repeated within aburst 66402 that has a finite duration or width 66403. The duration66403 is such that a duty cycle which can be defined as a ratio of burstduration to signal period is between about 1 to about 10⁻⁵. Preferablypseudo rectangular 10 microsecond pulses for pulse 66401 applied in aburst 66402 for about 10 to about 50 msec having a modified 1/famplitude envelope 66404 and with a finite duration 66403 correspondingto a burst period of between about 0.1 and about 10 seconds areutilized.

FIG. 67 illustrates an embodiment of an apparatus according to thepresent invention. A garment 67501 such as a vest is constructed out ofmaterials that are lightweight and portable such as nylon but othermaterials can be used. A miniature control circuit 67502 is coupled to agenerating device such as an electrical coil 67503. Preferably theminiature control circuit 67502 and the electrical coil 67503 areconstructed in a manner as described above in reference to FIG. 64. Theminiature control circuit and the electrical coil can be connected witha connecting means such as a wire 57504. The connection can also bedirect or wireless. The electrical coil 57503 is integrated into thegarment 57501 such that when a user wears the garment 57501, theelectrical coil is positioned near a lung or both lungs of the user. Anadvantage of the present invention is that its ultra lightweight coilsand miniaturized circuitry allow for the garment 57501 to be completelyself-contained, portable, and lightweight. An additionally advantageousresult of the present invention is that the garment 57501 can beconstructed to be inconspicuous when worn and can be worn as an outergarment such as a shirt or under other garments, so that only the userwill know that the garment 57501 is being worn and treatment is beingapplied. Use with common physical therapy treatment modalities, and atany respiratory location for which tissue growth, pain relief, andtissue and organ healing is easily obtained. An advantageous result ofapplication of the present invention is that tissue growth, repair, andmaintenance can be accomplished and enhanced anywhere and at anytime.Yet another advantageous result of application of the present inventionis that growth, repair, and maintenance of molecules, cells, tissues,and organs can be accomplished and enhanced anywhere and at anytime.Another embodiment according to the present invention delivers PEMF forapplication to respiratory tissue that is infected with diseases such assarcoidosis, granulomatous pneumonitis, pulmonary fibrosis, and “WorldTrade Center Cough.”

It is further intended that any other embodiments of the presentinvention that result from any changes in application or method of useor operation, method of manufacture, shape, size or material which arenot specified within the detailed written description or illustrationsand drawings contained herein, yet are considered apparent or obvious toone skilled in the art, are within the scope of the present invention.

The process of the invention will now be described with reference to thefollowing illustrative examples.

Example 1

The Power SNR approach for PMF signal configuration has been testedexperimentally on calcium dependent myosin phosphorylation in a standardenzyme assay. The cell-free reaction mixture was chosen forphosphorylation rate to be linear in time for several minutes, and forsub-saturation Ca²⁺ concentration. This opens the biological window forCa²⁺/CaM to be EMF-sensitive. This system is not responsive to PMF atlevels utilized in this study if Ca²⁺ is at saturation levels withrespect to CaM, and reaction is not slowed to a minute time range.Experiments were performed using myosin light chain (“MLC”) and myosinlight chain kinase (“MLCK”) isolated from turkey gizzard. A reactionmixture consisted of a basic solution containing 40 mM Hepes buffer, pH7.0; 0.5 mM magnesium acetate; 1 mg/ml bovine serum albumin, 0.1% (w/v)Tween 80; and 1 mM EGTA12. Free Ca²⁺ was varied in the 1-7 μM range.Once Ca²⁺ buffering was established, freshly prepared 70 nM CaM, 160 nMMLC and 2 nM MLCK were added to the basic solution to form a finalreaction mixture. The low MLC/MLCK ratio allowed linear time behavior inthe minute time range. This provided reproducible enzyme activities andminimized pipetting time errors.

The reaction mixture was freshly prepared daily for each series ofexperiments and was aliquoted in 100 μL portions into 1.5 ml Eppendorftubes. All Eppendorf tubes containing reaction mixture were kept at 0°C. then transferred to a specially designed water bath maintained at37±0.1° C. by constant perfusion of water prewarmed by passage through aFisher Scientific model 900 heat exchanger. Temperature was monitoredwith a thermistor probe such as a Cole-Parmer model 8110-20, immersed inone Eppendorf tube during all experiments. Reaction was initiated with2.5 μM 32P ATP, and was stopped with Laemmli Sample Buffer solutioncontaining 30 μM EDTA. A minimum of five blank samples were counted ineach experiment. Blanks comprised a total assay mixture minus one of theactive components Ca²⁺, CaM, MLC or MLCK. Experiments for which blankcounts were higher than 300 cpm were rejected. Phosphorylation wasallowed to proceed for 5 min and was evaluated by counting 32Pincorporated in MLC using a TM Analytic model 5303 Mark V liquidscintillation counter.

The signal comprised repetitive bursts of a high frequency waveform.Amplitude was maintained constant at 0.2 G and repetition rate was 1burst/sec for all exposures. Burst duration varied from 65 μsec to 1000μsec based upon projections of Power SNR analysis which showed thatoptimal Power SNR would be achieved as burst duration approached 500μsec. The results are shown in FIG. 68 wherein burst width 68601 in msecis plotted on the x-axis and Myosin Phosphorylation 68602 astreated/sham is plotted on the y-axis. It can be seen that the PMFeffect on Ca²⁺ binding to CaM approaches its maximum at approximately500 μsec, just as illustrated by the Power SNR model.

These results confirm that a PMF signal, configured according to anembodiment of the present invention, would maximally increase myosinphosphorylation for burst durations sufficient to achieve optimal PowerSNR for a given magnetic field amplitude.

Example 2

According to an embodiment of the present invention use of a Power SNRmodel was further verified in an in vivo wound repair model. A rat woundmodel has been well characterized both biomechanically andbiochemically, and was used in this study. Healthy, young adult maleSprague Dawley rats weighing more than 300 grams were utilized.

The animals were anesthetized with an intraperitoneal dose of Ketamine75 mg/kg and Medetomidine 0.5 mg/kg. After adequate anesthesia had beenachieved, the dorsum was shaved, prepped with a dilute betadine/alcoholsolution, and draped using sterile technique. Using a #10 scalpel, an8-cm linear incision was performed through the skin down to the fasciaon the dorsum of each rat. The wound edges were bluntly dissected tobreak any remaining dermal fibers, leaving an open wound approximately 4cm in diameter. Hemostasis was obtained with applied pressure to avoidany damage to the skin edges. The skin edges were then closed with a 4-0Ethilon running suture. Post-operatively, the animals receivedBuprenorphine 0.1-0.5 mg/kg, intraperitoneal. They were placed inindividual cages and received food and water ad libitum.

PMF exposure comprised two pulsed radio frequency waveforms. The firstwas a standard clinical PRF signal comprising a 65 μsec burst of 27.12MHz sinusoidal waves at 1 Gauss amplitude and repeating at 600bursts/sec. The second was a PRF signal reconfigured according to anembodiment of the present invention. For this signal burst duration wasincreased to 2000 μsec and the amplitude and repetition rate werereduced to 0.2 G and 5 bursts/sec respectively. PRF was applied for 30minutes twice daily.

Tensile strength was performed immediately after wound excision. Two 1cm width strips of skin were transected perpendicular to the scar fromeach sample and used to measure the tensile strength in kg/mm2. Thestrips were excised from the same area in each rat to assure consistencyof measurement. The strips were then mounted on a tensiometer. Thestrips were loaded at 10 mm/min and the maximum force generated beforethe wound pulled apart was recorded. The final tensile strength forcomparison was determined by taking the average of the maximum load inkilograms per mm2 of the two strips from the same wound.

The results showed average tensile strength for the 65 μsec 1 Gauss PRFsignal was 19.3±4.3 kg/mm2 for the exposed group versus 13.0±3.5 kg/mm2for the control group (p<0.01), which is a 48% increase. In contrast,the average tensile strength for the 2000 μsec 0.2 Gauss PRF signal,configured according to an embodiment of the present invention using aPower SNR model was 21.2±5.6 kg/mm2 for the treated group versus13.7±4.1 kg/mm2 (p<0.01) for the control group, which is a 54% increase.The results for the two signals were not significantly different fromeach other.

These results demonstrate that an embodiment of the present inventionallowed a new PRF signal to be configured that could be produced withsignificantly lower power. The PRF signal configured according to anembodiment of the present invention, accelerated wound repair in the ratmodel in a low power manner versus that for a clinical PRF signal whichaccelerated wound repair but required more than two orders of magnitudemore power to produce.

Example 3

This example illustrates the effects of PMF stimulation of a T-cellreceptor with cell arrest and thus behave as normal T-lymphocytesstimulated by antigens at the T-cell receptor such as anti-CD3.

In bone healing, results have shown that both 60 Hz and PEMF fieldsdecrease DNA synthesis of Jurkat cells, as is expected since PMFinteracts with the T-cell receptor in the absence of a costimulatorysignal. This result is consistent with an anti-inflammatory response, ashas been observed in clinical applications of PMF stimuli. The PEMFsignal is more effective. A dismetry analysis performed according to anembodiment of the present invention demonstrates why both signals areeffective and why PEMF signals have a greater effect than 60 Hz signalson Jurkat cells in the most EMF-sensitive growth stage.

Comparison of dosimetry from the two signals employed involvesevaluation of the ratio of the Power spectrum of the thermal noisevoltage that is Power SNR, to that of the induced voltage at theEMF-sensitive target pathway structure. The target pathway structureused is ion binding at receptor sites on Jurkat cells suspended in 2 mmof culture medium. The average peak electric field at the binding sitefrom a PEMF signal comprising 5 msec burst of 200 μsec pulses repeatingat 15/sec was 1 mV/cm, while for a 60 Hz signal the average peakelectric field was 100 μV/cm.

FIG. 69 is a graph of results wherein Induced Field Frequency 69701 inHz is shown on the x-axis and Power SNR 69702 is shown on the y-axis.FIG. 69 illustrates that both signals have sufficient Power spectrumthat is Power SNR≧1, to be detected within a frequency range of bindingkinetics. However, maximum Power SNR for the PEMF signal issignificantly higher than that of the 60 Hz signal. This is due to aPEMF signal having many frequency components falling within a bandpassof the target pathway structure. The single frequency component of a 60Hz signal lies at the mid-point of the bandpass of a target pathwaystructure. The Power SNR calculation that was used in this example isdependent upon τ_(ion) which is obtained from the rate constant for ionbinding. Had this calculation been performed a priori it would haveconcluded that both signals satisfied basic detectability requirementsand could modulate an EMF-sensitive ion binding pathway at the start ofa regulatory cascade for DNA synthesis in these cells. The previousexamples illustrate that utilizing the rate constant for Ca/CaM bindingcould lead to successful projections for bioeffective EMF signals in avariety of systems.

While the apparatus and method have been described in terms of what arepresently considered to be the most practical and preferred embodiments,it is to be understood that the disclosure need not be limited to thedisclosed embodiments. It is intended to cover various modifications andsimilar arrangements included within the spirit and scope of the claims,the scope of which should be accorded the broadest interpretation so asto encompass all such modifications and similar structures. The presentdisclosure includes any and all embodiments of the following claims.

Part 10

Induced time-varying currents from PEMF or PRF devices flow in a fibrouscapsule formation and capsular contracture target pathway structure suchas a molecule, cell, tissue, and organ, and it is these currents thatare a stimulus to which cells and tissues can react in a physiologicallymeaningful manner. The electrical properties of a fibrous capsuleformation and capsular contracture target pathway structure affectlevels and distributions of induced current. Molecules, cells, tissue,and organs are all in an induced current pathway such as cells in a gapjunction contact. Ion or ligand interactions at binding sites onmacromolecules that may reside on a membrane surface are voltagedependent chemical processes, that is electrochemical, that can respondto an induced electromagnetic field (“E”). Induced current arrives atthese sites via a surrounding ionic medium. The presence of cells in acurrent pathway causes an induced current (“J”) to decay more rapidlywith time (“J(t)”). This is due to an added electrical impedance ofcells from membrane capacitance and ion binding time constants ofbinding and other voltage sensitive membrane processes such as membranetransport. Knowledge of ion binding time constants allows SNR to beevaluated for any EMF signal configuration. Preferably ion binding timeconstants in the range of about 1 to about 100 msec are used.

Equivalent electrical circuit models representing various membrane andcharged interface configurations have been derived. For example, inCalcium (“Ca²⁺”) binding, the change in concentration of bound Ca²⁺ at abinding site due to induced E may be described in a frequency domain byan impedance expression such as:

${Z_{b}\; (\omega)} = {R_{ion} + \frac{1}{i\; \omega \; C_{ion}}}$

which has the form of a series resistance-capacitance electricalequivalent circuit. Where ω is angular frequency defined as 2πf, where fis frequency, i=−1^(1/2), Z_(b)(ω) is the binding impedance, and R_(ion)and C_(ion) are equivalent binding resistance and capacitance of an ionbinding pathway. The value of the equivalent binding time constant,τ_(ion)=R_(ion)C_(ion), is related to a ion binding rate constant,k_(b), via τ_(ion)=R_(ion)C_(ion)=1/k_(b). Thus, the characteristic timeconstant of this pathway is determined by ion binding kinetics.

Induced E from a PEMF or PRF signal can cause current to flow into anion binding pathway and affect the number of Ca²⁺ ions bound per unittime. An electrical equivalent of this is a change in voltage across theequivalent binding capacitance C_(ion), which is a direct measure of thechange in electrical charge stored by C_(ion). Electrical charge isdirectly proportional to a surface concentration of Ca²⁺ ions in thebinding site, that is storage of charge is equivalent to storage of ionsor other charged species on cell surfaces and junctions. Electricalimpedance measurements, as well as direct kinetic analyses of bindingrate constants, provide values for time constants necessary forconfiguration of a PMF waveform to match a bandpass of target pathwaystructures. This allows for a required range of frequencies for anygiven induced E waveform for optimal coupling to target impedance, suchas bandpass.

Ion binding to regulatory molecules is a frequent EMF target, forexample Ca²⁺ binding to calmodulin (“CaM”). Use of this pathway is basedupon acceleration of wound repair, for example bone repair, thatinvolves modulation of growth factors released in various stages ofrepair. Growth factors such as platelet derived growth factor (“PDGF”),fibroblast growth factor (“FGF”), and epidermal growth factor (“EGF”)are all involved at an appropriate stage of healing. Angiogenesis isalso integral to wound repair and modulated by PMF. All of these factorsare Ca/CaM-dependent.

Utilizing a Ca/CaM pathway a waveform can be configured for whichinduced power is sufficiently above background thermal noise power.Under correct physiological conditions, this waveform can have aphysiologically significant bioeffect.

Application of a Power SNR model to Ca/CaM requires knowledge ofelectrical equivalents of Ca²⁺ binding kinetics at CaM. Within firstorder binding kinetics, changes in concentration of bound Ca²⁺ at CaMbinding sites over time may be characterized in a frequency domain by anequivalent binding time constant, τ_(ion)=R_(ion)C_(ion), where R_(ion)and C_(ion) are equivalent binding resistance and capacitance of the ionbinding pathway. τ_(ion) is related to a ion binding rate constant,k_(b), via τ_(ion)=R_(ion)C_(ion)=1/k_(b). Published values for k_(b)can then be employed in a cell array model to evaluate SNR by comparingvoltage induced by a PRF signal to thermal fluctuations in voltage at aCaM binding site. Employing numerical values for PMF response, such asV_(max)=6.5×10⁻⁷ sec⁻¹, =2.50/1, K_(D)=30 μM, [Ca²⁺CaM]=K_(D)(+[CaM]),yields k_(b)=665 sec⁻¹ (τ_(ion)=1.5 msec). Such a value for τ_(ion) canbe employed in an electrical equivalent circuit for ion binding whilepower SNR analysis can be performed for any waveform structure.

According to an embodiment of the present invention a mathematical modelcan be configured to assimilate that thermal noise is present in allvoltage dependent processes and represents a minimum thresholdrequirement to establish adequate SNR. Power spectral density, S_(n)(ω),of thermal noise can be expressed as:

S _(n)(ω)=4kT Re[Z _(M)(x,ω)]

where Z_(M)(x, ω) is electrical impedance of a target pathway structure,x is a dimension of a target pathway structure and Re denotes a realpart of impedance of a target pathway structure. Z_(M)(x, ω) can beexpressed as:

${Z_{M}( {x,\omega} )} = {\lbrack \frac{R_{e} + R_{i} + R_{g}}{y} \rbrack \tanh \; ( {y\; x} )}$

This equation clearly shows that electrical impedance of the targetpathway structure, and contributions from extracellular fluid resistance(“R_(e)”), intracellular fluid resistance (“R_(i)”) and intermembraneresistance (“R_(g)”) which are electrically connected to a targetpathway structure, all contribute to noise filtering.

A typical approach to evaluation of SNR uses a single value of a rootmean square (RMS) noise voltage. This is calculated by taking a squareroot of an integration of S_(n)(ω)=4 kT Re [Z_(M)(x, ω)] over allfrequencies relevant to either complete membrane response, or tobandwidth of a target pathway structure. SNR can be expressed by aratio:

${SNR} = \frac{{V_{M}(\omega)}}{RMS}$

where |V_(M)(ω)| is maximum amplitude of voltage at each frequency asdelivered by a chosen waveform to the target pathway structure.

An embodiment according to the present invention comprises a pulse burstenvelope having a high spectral density, so that the effect of therapyupon the relevant dielectric pathways, such as, cellular membranereceptors, ion binding to cellular enzymes and general transmembranepotential changes, is enhanced. Accordingly by increasing a number offrequency components transmitted to relevant cellular pathways, a largerange of biophysical phenomena, such as modulating growth factor andcytokine release and ion binding at regulatory molecules, applicable toknown tissue growth mechanisms is accessible. According to an embodimentof the present invention applying a random, or other high spectraldensity envelope, to a pulse burst envelope of mono-polar or bi-polarrectangular or sinusoidal pulses inducing peak electric fields betweenabout 10⁻⁸ and about 100 mV/cm, produces a greater effect on biologicalhealing processes applicable to both soft and hard tissues.

An embodiment according to the present invention comprises anelectromagnetic signal having a pulse burst envelope of spectral densityto efficiently couple to physiologically relevant dielectric pathways,such as cellular membrane receptors, ion binding to cellular enzymes,and general transmembrane potential changes. The use of a burst durationwhich is generally below 100 microseconds for each PRF burst, limits thefrequency components that could couple to the relevant dielectricpathways in cells and tissue. An embodiment according to the presentinvention increases the number of frequency components transmitted torelevant cellular pathways whereby access to a larger range ofbiophysical phenomena applicable to known healing mechanisms, includingenhanced second messenger release, enzyme activity and growth factor andcytokine release can be achieved. By increasing burst duration andapplying a random, or other envelope, to the pulse burst envelope ofmono-polar or bi-polar rectangular or sinusoidal pulses which inducepeak electric fields between 10⁻⁸ and 100 mV/cm, a more efficient andgreater effect can be achieved on biological healing processesapplicable to both soft and hard tissues in humans, animals and plants.

Another embodiment according to the present invention comprises knowncellular responses to weak external stimuli such as heat, light, sound,ultrasound and electromagnetic fields. Cellular responses to suchstimuli result in the production of protective proteins, for example,heat shock proteins, which enhance the ability of the cell, tissue,organ to withstand and respond to such external stimuli. Electromagneticfields configured according to an embodiment of the present inventionenhance the release of such compounds thus advantageously providing animproved means to enhance prophylactic protection and wellness of livingorganisms. After implant surgery there can be physiological deficienciessuch as capsular contraction and excessive fibrous capsule formationstates that can have a lasting and deleterious effect on an individual'swell being and on the proper functioning of an implanted device. Thosephysiological deficiencies and states can be positively affected on anon-invasive basis by the therapeutic application of waveformsconfigured according to an embodiment of the present invention. Inaddition, electromagnetic waveforms configured according to anembodiment of the present invention can have a prophylactic effect on animplant area whereby formation of excessive fibrous tissue may beprevented.

The present invention relates to a therapeutically beneficial method ofand apparatus for non-invasive pulsed electromagnetic treatment forenhanced condition, repair and growth of living tissue in animals,humans and plants. This beneficial method operates to selectively changethe bioelectromagnetic environment associated with the cellular andtissue environment through the use of electromagnetic means such as PRFgenerators and applicator heads. More particularly use ofelectromagnetic means includes the provision of a flux path to aselectable body region, of a succession of EMF pulses having a minimumwidth characteristic of at least 0.01 microseconds in a pulse burstenvelope having between 1 and 100,000 pulses per burst, in which avoltage amplitude envelope of said pulse burst is defined by a randomlyvarying parameter. Further, the repetition rate of such pulse bursts mayvary from 0.01 to 10,000 Hz. Additionally a mathematically-definableparameter can be employed in lieu of said random amplitude envelope ofthe pulse bursts.

According to an embodiment of the present invention, by applying arandom, or other high spectral density envelope, to a pulse burstenvelope of mono-polar or bi-polar rectangular or sinusoidal pulseswhich induce peak electric fields between 10⁻⁸ and 100 millivolts percentimeter (mV/cm), a more efficient and greater effect can be achievedon biological healing processes applicable to both soft and hard tissuesin humans, animals and plants. A pulse burst envelope of higher spectraldensity can advantageously and efficiently couple to physiologicallyrelevant dielectric pathways, such as, cellular membrane receptors, ionbinding to cellular enzymes, and general transmembrane potential changesthereby modulating angiogenesis and neovascularization.

An embodiment according to the present invention utilizes a Power Signalto Noise Ratio (“Power SNR”) approach to configure bioeffectivewaveforms and incorporates miniaturized circuitry and lightweightflexible coils. This advantageously allows a device that utilizes aPower SNR approach, miniaturized circuitry, and lightweight flexiblecoils, to be completely portable and if desired to be constructed asdisposable and if further desired to be constructed as implantable. Thelightweight flexible coils can be an integral portion of a positioningdevice such as surgical dressings, wound dressings, pads, seat cushions,mattress pads, wheelchairs, chairs, and any other garment and structurejuxtaposed to living tissue and cells. By advantageously integrating acoil into a positioning device therapeutic treatment can be provided toliving tissue and cells in an inconspicuous and convenient manner.

Specifically, broad spectral density bursts of electromagneticwaveforms, configured to achieve maximum signal power within a bandpassof a biological target, are selectively applied to fibrous capsuleformation and capsular contracture target pathway structures such asliving organs, tissues, cells and molecules that are associated withexcessive fibrous capsule formation and capsular contracture. Waveformsare selected using a novel amplitude/power comparison with that ofthermal noise in a fibrous capsule formation and capsular contracturetarget pathway structure. Signals comprise bursts of at least one ofsinusoidal, rectangular, chaotic and random wave shapes have frequencycontent in a range of 0.01 Hz to 100 MHz at 1 to 100,000 bursts persecond, with a burst duration from 0.01 to 100 milliseconds, and a burstrepetition rate from 0.01 to 1000 bursts/second. Peak signal amplitudeat a fibrous capsule formation and capsular contracture target pathwaystructure such as tissue, lies in a range of 1 μV/cm to 100 mV/cm. Eachsignal burst envelope may be a random function providing a means toaccommodate different electromagnetic characteristics of healing tissue.Preferably the present invention comprises a 20 millisecond pulse burst,repeating at 1 to 10 burst/second and comprising 0.5 to 200 microsecondsymmetrical or asymmetrical pulses repeating at 10⁻⁵ to 100 kilohertzwithin the burst. The burst envelope can be modified 1/f function or anyarbitrary function and can be applied at random repetition rates. Fixedrepetition rates can also be used between about 0.1 Hz and about 1000Hz. An induced electric field from about 10⁻⁸ mV/cm to about 100 mV/cmis generated. Another embodiment according to the present inventioncomprises a 4 millisecond of high frequency sinusoidal waves, such as27.12 MHz, repeating at 1 to 100 bursts per second. An induced electricfield from about 10⁻⁸ mV/cm to about 100 mV/cm is generated. Resultingwaveforms can be delivered via inductive or capacitive coupling for 1 to30 minute treatment sessions delivered according to predefined regimesby which PEMF treatment may be applied for 1 to 12 daily sessions,repeated daily. The treatment regimens for any waveform configuredaccording to the instant invention may be fully automated. The number ofdaily treatments may be programmed to vary on a daily basis according toany predefined protocol.

According to yet another embodiment of the present invention by applyinga high spectral density voltage envelope as a modulating or pulse-burstdefining parameter, power requirements for such amplitude modulatedpulse bursts can be significantly lower than that of an unmodulated orcontinuous pulse burst containing pulses within a similar carrierfrequency range. This is due to a substantial reduction in duty cyclewithin repetitive burst trains brought about by imposition of anirregular amplitude and preferably a random amplitude onto what wouldotherwise be a substantially uniform pulse burst envelope. Accordingly,the dual advantages, of enhanced transmitted dosimetry to the relevantdielectric pathways and of decreased power requirement are achieved.

Referring to FIG. 70 wherein FIG. 70 is a flow diagram of a method forgenerating electromagnetic signals to be coupled to a fibrous capsuleformation and capsular contracture target pathway structure according toan embodiment of the present invention, a fibrous capsule formation andcapsular contracture target pathway structure such as ions and ligands,is identified. Establishing a baseline background activity such asbaseline thermal fluctuations in voltage and electrical impedance, atthe fibrous capsule formation and capsular contracture target pathwaystructure by determining a state of at least one of a cell and a tissueat the fibrous capsule formation and capsular contracture target pathwaystructure, wherein the state is at least one of resting, growing,replacing, and responding to injury. (STEP 70101) The state of the atleast one of a cell and a tissue is determined by its response to injuryor insult. Configuring at least one waveform to have sufficient signalto noise ratio to modulate at least one of ion and ligand interactionswhereby the at least one of ion and ligand interactions are detectablein the fibrous capsule formation and capsular contracture target pathwaystructure above the established baseline thermal fluctuations in voltageand electrical impedance. The EMF signal can be generated by using atleast one waveform configured by applying a mathematical model such asan equation, formula, or function having at least one waveform parameterthat satisfies an SNR or Power SNR mathematical model of at least about0.2, to modulate at least one of ion and ligand interactions whereby theat least one of ion and ligand interactions are detectable in a fibrouscapsule formation and capsular contracture target pathway structureabove baseline thermal fluctuations in voltage and electrical impedanceat the fibrous capsule formation and capsular contracture target pathwaystructure, wherein the signal to noise ratio is evaluated by calculatinga frequency response of the impedance of the target path structuredivided by a calculated frequency response of baseline thermalfluctuations in voltage across the target path structure (STEP 70102).Repetitively generating an electromagnetic signal from the configured atleast one waveform (STEP 70103). Coupling the electromagnetic signal tothe fibrous capsule formation and capsular contracture target pathwaystructure using a coupling device (STEP 70104). The generatedelectromagnetic signals can be coupled for therapeutic and prophylacticpurposes. The coupling enhances a stimulus that cells and tissues reactto in a physiological meaningful manner for example, an increase inangiogenesis, neovascularization and vascularogenesis or otherphysiological effects related to the improvement of excessive fibroustissue or capsular contracture. Application of electromagnetic signalsusing an embodiment according to the present invention is extremely safeand efficient since the application of electromagnetic signalsconfigured according to the present invention is non-invasive andathermal.

In the present invention, a generated electromagnetic signal iscomprised of a burst of arbitrary waveforms having at least one waveformparameter that includes a plurality of frequency components ranging fromabout 0.01 Hz to about 100 MHz wherein the plurality of frequencycomponents satisfies a Power SNR model. A repetitive electromagneticsignal can be generated for example inductively or capacitively, fromthe configured at least one waveform. The electromagnetic signal iscoupled to a fibrous capsule formation and capsular contracture targetpathway structure such as ions and ligands by output of a couplingdevice such as an electrode or an inductor, placed in close proximity tothe fibrous capsule formation and capsular contracture target pathwaystructure using a positioning device. The coupling enhances modulationof binding of ions and ligands to regulatory molecules, tissues, cells,and organs. According to an embodiment of the present invention EMFsignals configured using SNR analysis to match the bandpass of a secondmessenger whereby the EMF signals can act as a first messenger tomodulate biochemical cascades such as production of cytokines, NitricOxide, Nitric Oxide Synthase and growth factors that are related totissue growth and repair. A detectable E field amplitude is producedwithin a frequency response of Ca²⁺ binding.

FIG. 71 illustrates an embodiment of an apparatus according to thepresent invention. The apparatus is constructed to be self-contained,lightweight, and portable. A miniature control circuit 71201 isconnected to a generating device such as an electrical coil 71202. Theminiature control circuit 71201 is constructed in a manner that appliesa mathematical model that is used to configure waveforms. The configuredwaveforms have to satisfy a Power SNR model so that for a given andknown fibrous capsule formation and capsular contracture target pathwaystructure, it is possible to choose waveform parameters that satisfy afrequency response of the fibrous capsule formation and capsularcontracture target pathway structure and Power SNR of at least about 0.2to modulate at least one of ion and ligand interactions whereby the atleast one of ion and ligand interactions are detectable in a fibrouscapsule formation and capsular contracture target pathway structureabove baseline thermal fluctuations in voltage and electrical impedanceat the fibrous capsule formation and capsular contracture target pathwaystructure, wherein the signal to noise ratio is evaluated by calculatinga frequency response of the impedance of the target path structuredivided by a calculated frequency response of baseline thermalfluctuations in voltage across the target path structure. An embodimentaccording to the present invention applies a mathematical model toinduce a time-varying magnetic field and a time-varying electric fieldin a fibrous capsule formation and capsular contracture target pathwaystructure such as ions and ligands, comprising about 0.001 to about 100msec bursts of about 1 to about 100 microsecond rectangular pulses,having a burst duration of about 0.01 to 100,000 microseconds andrepeating at about 0.1 to about 100 pulses per second. Peak amplitude ofthe induced electric field is between about 1 uV/cm and about 100 mV/cm,that can be constant or varied according to a mathematical function, forexample a modified 1/f function where f=frequency. A waveform configuredusing an embodiment according to the present invention may be applied toa fibrous capsule formation and capsular contracture target pathwaystructure such as ions and ligands, preferably for a total exposure timeof under 1 minute to 240 minutes daily. However other exposure times canbe used. Waveforms configured by the miniature control circuit 71201 aredirected to a generating device 71202 such as electrical coils.Preferably, the generating device 71202 is a conformable coil forexample pliable, comprising one or more turns of electrically conductingwire in a generally circular or oval shape however other shapes can beused. The generating device 71202 delivers a pulsing magnetic fieldconfigured according to a mathematical model that can be used to providetreatment to a fibrous capsule formation and capsular contracture targetpathway structure such as mammary tissue. The miniature control circuitapplies a pulsing magnetic field for a prescribed time and canautomatically repeat applying the pulsing magnetic field for as manyapplications as are needed in a given time period, for example 12 timesa day. The miniature control circuit can be configured to beprogrammable applying pulsing magnetic fields for any time repetitionsequence. An embodiment according to the present invention can bepositioned to treat fibrous capsule tissue by being incorporated with apositioning device such as a bandage, a vest, a brassiere, or ananatomical support thereby making the unit self-contained. Coupling apulsing magnetic field to a fibrous capsule formation and capsularcontracture target pathway structure such as ions and ligands,therapeutically and prophylactically reduces inflammation therebyreducing pain and promotes healing in treatment areas. When electricalcoils are used as the generating device 71202, the electrical coils canbe powered with a time varying magnetic field that induces a timevarying electric field in a fibrous capsule formation and capsularcontracture target pathway structure according to Faraday's law. Anelectromagnetic signal generated by the generating device 202 can alsobe applied using electrochemical coupling, wherein electrodes are indirect contact with skin or another outer electrically conductiveboundary of a fibrous capsule formation and capsular contracture targetpathway structure. Yet in another embodiment according to the presentinvention, the electromagnetic signal generated by the generating device202 can also be applied using electrostatic coupling wherein an air gapexists between a generating device 202 such as an electrode and afibrous capsule formation and capsular contracture target pathwaystructure such as ions and ligands. An advantage of the presentinvention is that its ultra lightweight coils and miniaturized circuitryallow for use with common physical therapy treatment modalities, and atany location for which tissue growth, pain relief, and tissue and organhealing is desired. An advantageous result of application of the presentinvention is that tissue growth, repair, and maintenance can beaccomplished and enhanced anywhere and at anytime. Yet anotheradvantageous result of application of the present invention is thatgrowth, repair, and maintenance of molecules, cells, tissues, and organscan be accomplished and enhanced anywhere and at anytime. Anotherembodiment according to the present invention delivers PEMF forapplication to capsular contracture and excessive fibrous capsule tissuethat resulted from implant surgery such as breast augmentation.

FIG. 72 depicts a block diagram of an embodiment according to thepresent invention of a miniature control circuit 72300. The miniaturecontrol circuit 72300 produces waveforms that drive a generating devicesuch as wire coils described above in FIG. 71. The miniature controlcircuit can be activated by any activation means such as an on/offswitch. The miniature control circuit 72300 has a power source such as alithium battery 72301. Preferably the power source has an output voltageof 3.3 V but other voltages can be used. In another embodiment accordingto the present invention the power source can be an external powersource such as an electric current outlet such as an AC/DC outlet,coupled to the present invention for example by a plug and wire. Aswitching power supply 72302 controls voltage to a micro-controller72303. Preferably the micro-controller 72303 uses an 8 bit 4 MHzmicro-controller 72303 but other bit MHz combination micro-controllersmay be used. The switching power supply 72302 also delivers current tostorage capacitors 72304. Preferably the storage capacitors 72304 havinga 220 uF output but other outputs can be used. The storage capacitors72304 allow high frequency pulses to be delivered to a coupling devicesuch as inductors (Not Shown). The micro-controller 72303 also controlsa pulse shaper 72305 and a pulse phase timing control 72306. The pulseshaper 72305 and pulse phase timing control 72306 determine pulse shape,burst width, burst envelope shape, and burst repetition rate. In anembodiment according to the present invention the pulse shaper 72305 andphase timing control 72306 are configured such that the waveformsconfigured are detectable above background activity at a fibrous capsuleformation and capsular contracture target pathway structure bysatisfying at least one of a SNR and Power SNR mathematical model. Anintegral waveform generator, such as a sine wave or arbitrary numbergenerator can also be incorporated to provide specific waveforms. Avoltage level conversion sub-circuit 72307 controls an induced fielddelivered to a fibrous capsule formation and capsular contracture targetpathway structure. A switching Hexfet 72308 allows pulses of randomizedamplitude to be delivered to output 72309 that routes a waveform to atleast one coupling device such as an inductor. The micro-controller72303 can also control total exposure time of a single treatment of afibrous capsule formation and capsular contracture target pathwaystructure such as a molecule, cell, tissue, and organ. The miniaturecontrol circuit 72300 can be constructed to be programmable and apply apulsing magnetic field for a prescribed time and to automatically repeatapplying the pulsing magnetic field for as many applications as areneeded in a given time period, for example 10 times a day. Preferablytreatments times of about 1 minutes to about 30 minutes are used.

Referring to FIG. 73 an embodiment according to the present invention ofa waveform 73400 is illustrated. A pulse 73401 is repeated within aburst 73402 that has a finite duration 73403 alternatively referred toas width 73403. The duration 73403 is such that a duty cycle which canbe defined as a ratio of burst duration to signal period is betweenabout 1 to about 10⁻⁵. Preferably pseudo rectangular 10 microsecondpulses for pulse 73401 applied in a burst 73402 for about 10 to about 50msec having a modified 1/f amplitude envelope 73404 and with a finiteduration 73403 corresponding to a burst period of between about 0.1 andabout 10 seconds are utilized.

FIG. 74 illustrates an embodiment of an apparatus according to thepresent invention. A garment 74501 such as a brassiere is constructedout of materials that are lightweight and portable such as nylon butother materials can be used. A miniature control circuit 74502 iscoupled to a generating device such as an electrical coil 74503.Preferably the miniature control circuit 74502 and the electrical coil74503 are constructed in a manner as described above in reference toFIG. 71. The miniature control circuit and the electrical coil can beconnected with a connecting means such as a wire 74504. The connectioncan also be direct or wireless. The electrical coil 74503 is integratedinto the garment 74501 such that when a user wears the garment 74501,the electrical coil is positioned near an excessive fibrous capsuleformation location or capsular contracture location of the user. Anadvantage of the present invention is that its ultra lightweight coilsand miniaturized circuitry allow for the garment 74501 to be completelyself-contained, portable, and lightweight. An additionally advantageousresult of the present invention is that the garment 74501 can beconstructed to be inconspicuous when worn and can be worn as an outergarment such as a shirt or under other garments, so that only the userwill know that the garment 74501 is being worn and treatment is beingapplied. Use with common physical therapy treatment modalities, and atany excessive fibrous capsule location or capsular contracture locationfor which pain relief, and tissue and organ healing is easily obtained.An advantageous result of application of the present invention is thattissue growth, repair, and maintenance can be accomplished and enhancedanywhere and at anytime. Yet another advantageous result of applicationof the present invention is that growth, repair, and maintenance ofmolecules, cells, tissues, and organs can be accomplished and enhancedanywhere and at anytime. Another embodiment according to the presentinvention delivers PEMF for application to fibrous capsules.

It is further intended that any other embodiments of the presentinvention that result from any changes in application or method of useor operation, method of manufacture, shape, size or material which arenot specified within the detailed written description or illustrationsand drawings contained herein, yet are considered apparent or obvious toone skilled in the art, are within the scope of the present invention.

The process of the invention will now be described with reference to thefollowing illustrative examples.

Example 1

The Power SNR approach for PMF signal configuration has been testedexperimentally on calcium dependent myosin phosphorylation in a standardenzyme assay. The cell-free reaction mixture was chosen forphosphorylation rate to be linear in time for several minutes, and forsub-saturation Ca²⁺ concentration. This opens the biological window forCa²⁺/CaM to be EMF-sensitive. This system is not responsive to PMF atlevels utilized in this study if Ca is at saturation levels with respectto CaM, and reaction is not slowed to a minute time range. Experimentswere performed using myosin light chain (“MLC”) and myosin light chainkinase (“MLCK”) isolated from turkey gizzard. A reaction mixtureconsisted of a basic solution containing 40 mM Hepes buffer, pH 7.0; 0.5mM magnesium acetate; 1 mg/ml bovine serum albumin, 0.1% (w/v) Tween80;and 1 mM EGTA12. Free Ca²⁺ was varied in the 1-7 μM range. Once Ca²⁺buffering was established, freshly prepared 70 nM CaM, 160 nM MLC and 2nM MLCK were added to the basic solution to form a final reactionmixture. The low MLC/MLCK ratio allowed linear time behavior in theminute time range. This provided reproducible enzyme activities andminimized pipetting time errors.

The reaction mixture was freshly prepared daily for each series ofexperiments and was aliquoted in 100 μL portions into 1.5 ml Eppendorftubes. All Eppendorf tubes containing reaction mixture were kept at 0°C. then transferred to a specially designed water bath maintained at37±0.1° C. by constant perfusion of water prewarmed by passage through aFisher Scientific model 900 heat exchanger. Temperature was monitoredwith a thermistor probe such as a Cole-Parmer model 8110-20, immersed inone Eppendorf tube during all experiments. Reaction was initiated with2.5 μM 32P ATP, and was stopped with Laemmli Sample Buffer solutioncontaining 30 μM EDTA. A minimum of five blank samples were counted ineach experiment. Blanks comprised a total assay mixture minus one of theactive components Ca²⁺, CaM, MLC or MLCK. Experiments for which blankcounts were higher than 300 cpm were rejected. Phosphorylation wasallowed to proceed for 5 min and was evaluated by counting 32Pincorporated in MLC using a TM Analytic model 5303 Mark V liquidscintillation counter.

The signal comprised repetitive bursts of a high frequency waveform.Amplitude was maintained constant at 0.2 G and repetition rate was 1burst/sec for all exposures. Burst duration varied from 65 μsec to 1000μsec based upon projections of Power SNR analysis which showed thatoptimal Power SNR would be achieved as burst duration approached 500μsec. The results are shown in FIG. 75 wherein burst width 75601 in msecis plotted on the x-axis and Myosin Phosphorylation 75602 astreated/sham is plotted on the y-axis. It can be seen that the PMFeffect on Ca²⁺ binding to CaM approaches its maximum at approximately500 μsec, just as illustrated by the Power SNR model.

These results confirm that a PMF signal, configured according to anembodiment of the present invention, would maximally increase myosinphosphorylation for burst durations sufficient to achieve optimal PowerSNR for a given magnetic field amplitude.

Example 2

According to an embodiment of the present invention use of a Power SNRmodel was further verified in an in vivo wound repair model. A rat woundmodel has been well characterized both biomechanically andbiochemically, and was used in this study. Healthy, young adult maleSprague Dawley rats weighing more than 300 grams were utilized.

The animals were anesthetized with an intraperitoneal dose of Ketamine75 mg/kg and Medetomidine 0.5 mg/kg. After adequate anesthesia had beenachieved, the dorsum was shaved, prepped with a dilute betadine/alcoholsolution, and draped using sterile technique. Using a #10 scalpel, an8-cm linear incision was performed through the skin down to the fasciaon the dorsum of each rat. The wound edges were bluntly dissected tobreak any remaining dermal fibers, leaving an open wound approximately 4cm in diameter. Hemostasis was obtained with applied pressure to avoidany damage to the skin edges. The skin edges were then closed with a 4-0Ethilon running suture. Post-operatively, the animals receivedBuprenorphine 0.1-0.5 mg/kg, intraperitoneal. They were placed inindividual cages and received food and water ad libitum.

PMF exposure comprised two pulsed radio frequency waveforms. The firstwas a standard clinical PRF signal comprising a 65 μsec burst of 27.12MHz sinusoidal waves at 1 Gauss amplitude and repeating at 600bursts/sec. The second was a PRF signal reconfigured according to anembodiment of the present invention. For this signal burst duration wasincreased to 2000 μsec and the amplitude and repetition rate werereduced to 0.2 G and 5 bursts/sec respectively. PRF was applied for 30minutes twice daily.

Tensile strength was performed immediately after wound excision. Two 1cm width strips of skin were transected perpendicular to the scar fromeach sample and used to measure the tensile strength in kg/mm2. Thestrips were excised from the same area in each rat to assure consistencyof measurement. The strips were then mounted on a tensiometer. Thestrips were loaded at 10 mm/min and the maximum force generated beforethe wound pulled apart was recorded. The final tensile strength forcomparison was determined by taking the average of the maximum load inkilograms per mm2 of the two strips from the same wound.

The results showed average tensile strength for the 65 μsec 1 Gauss PRFsignal was 19.3±4.3 kg/mm2 for the exposed group versus 13.0±3.5 kg/mm2for the control group (p<0.01), which is a 48% increase. In contrast,the average tensile strength for the 2000 μsec 0.2 Gauss PRF signal,configured according to an embodiment of the present invention using aPower SNR model was 21.2±5.6 kg/mm2 for the treated group versus13.7±4.1 kg/mm2 (p<0.01) for the control group, which is a 54% increase.The results for the two signals were not significantly different fromeach other.

These results demonstrate that an embodiment of the present inventionallowed a new PRF signal to be configured that could be produced withsignificantly lower power. The PRF signal configured according to anembodiment of the present invention, accelerated wound repair in the ratmodel in a low power manner versus that for a clinical PRF signal whichaccelerated wound repair but required more than two orders of magnitudemore power to produce.

Example 3

This example illustrates the effects of PMF stimulation of a T-cellreceptor with cell arrest and thus behave as normal T-lymphocytesstimulated by antigens at the T-cell receptor such as anti-CD3.

In bone healing, results have shown that both 60 Hz and PEMF fieldsdecrease DNA synthesis of Jurkat cells, as is expected since PMFinteracts with the T-cell receptor in the absence of a costimulatorysignal. This result is consistent with an anti-inflammatory response, ashas been observed in clinical applications of PMF stimuli. The PEMFsignal is more effective. A dosimetry analysis performed according to anembodiment of the present invention demonstrates why both signals areeffective and why PEMF signals have a greater effect than 60 Hz signalson Jurkat cells in the most EMF-sensitive growth stage.

Comparison of dosimetry from the two signals employed involvesevaluation of the ratio of the Power spectrum of the thermal noisevoltage that is Power SNR, to that of the induced voltage at theEMF-sensitive target pathway structure. The target pathway structureused is ion binding at receptor sites on Jurkat cells suspended in 2 mmof culture medium. The average peak electric field at the binding sitefrom a PEMF signal comprising 5 msec burst of 200 μsec pulses repeatingat 15/sec was 1 mV/cm, while for a 60 Hz signal the average peakelectric field was 100 μV/cm.

FIG. 76 is a graph of results wherein Induced Field Frequency 76701 inHz is shown on the x-axis and Power SNR 702 is shown on the y-axis. FIG.76 illustrates that both signals have sufficient Power spectrum that isPower SNR 1, to be detected within a frequency range of bindingkinetics. However, maximum Power SNR for the PEMF signal issignificantly higher than that of the 60 Hz signal. This is due to aPEMF signal having many frequency components falling within a bandpassof the target pathway structure. The single frequency component of a 60Hz signal lies at the mid-point of the bandpass of a target pathwaystructure. The Power SNR calculation that was used in this example isdependent upon τ_(ion) which is obtained from the rate constant for ionbinding. Had this calculation been performed a priori it would haveconcluded that both signals satisfied basic detectability requirementsand could modulate an EMF-sensitive ion binding pathway at the start ofa regulatory cascade for DNA synthesis in these cells. The previousexamples illustrate that utilizing the rate constant for Ca/CaM bindingcould lead to successful projections for bioeffective EMF signals in avariety of systems.

Example 4

In this example six patients who had developed capsular contractureafter receiving bilateral breast implants were treated with a specialsupport brassiere having embedded coils located in each cup and agenerator for each coil located in a special pocket in the strap aboveeach cup as described in FIG. 5 above. PEMF signals generated by theapparatus configured according to an embodiment of the present inventioncomprised a repetitive burst of radio frequency sinusoidal wavesconfigured according to an embodiment of the present invention. The PEMFsignal induced a peak electric field in a range of 1 to 10 mV/cm. Allpatients were provided a regimen that comprised six thirty minutesessions for days 1 to 3 post implant, four sessions for days 4 to 6post implant, and two sessions for all subsequent days. Clinicalevaluation demonstrated that by day 7 the fibrous capsule wassignificantly softer and patients reported significantly less pain anddiscomfort than prior to the treatment. Clinical evaluations at one andthree months post PEMF treatment revealed significant resolution of thefibrous capsule and its corresponding symptoms.

Part 11

Basal levels of intracellular Ca²⁺ are typically 50-100 nM, tightlymaintained by a number of physiological calcium buffers. It is generallyaccepted that transient elevations in cytosolic Ca²⁺ from externalstimuli as simple as changes in temperature and mechanical forces, or ascomplex as mechanical disruption of tissue, rapidly activate CaM, whichequally rapidly activates the cNOS enzymes, i.e., endothelial andneuronal NOS, or eNOS and nNOS, respectively. Studies have shown thatboth isoforms are inactive at basal intracellular levels of Ca²⁺,however, their activity increases with elevated Ca²⁺, reachinghalf-maximal activity at about 300 nM. Thus, nNOS and eNOS are regulatedby changes in intracellular Ca²⁺ concentrations within the physiologicalrange. In contrast, a third, inducible isoform of NOS (iNOS), which isupregulated during inflammation by macrophages and/or neutrophils,contains CaM that is tightly bound, even at low resting levels ofcytosolic Ca²⁺, and is not sensitive to intracellular Ca²⁺.

Once cNOS is activated by CaM it converts its substrate, L-arginine, tocitrulline, releasing one molecule of NO. As a gaseous free radical witha half-life of about 5 sec, NO diffuses locally through membranes andorganelles and acts on molecular targets at a distance up to about 200μm. The low transient concentrations of NO from cNOS can activatesoluble guanylyl cyclase (sGC), which catalyzes the synthesis of cyclicguanosine monophosphate (cGMP). The CaM/NO/cGMP signaling pathway is arapid response cascade which can modulate peripheral and cardiac bloodflow in response to normal physiologic demands, as well as toinflammation. This same pathway also modulates the release of cytokines,such as interleukin-1beta (IL-1β) and growth factors such as basicfibroblast growth factor (FGF-2) and vascular endothelial growth factor(VEGF) which have pleiotropic effects on cells involved in tissue repairand maintenance.

Following an injury, e.g., a bone fracture, torn rotator cuff, sprain,strain or surgical incision, repair commences with an inflammatory stageduring which the pro-inflammatory cytokine IL-1β is rapidly released.This, in turn, up-regulates iNOS, resulting in the production of largeamounts of NO in the wound bed. Continued exposure to NO leads to theinduction of cyclooxygenase-2 and increased synthesis of prostaglandinswhich also play a role in the inflammatory phase. While this process isa natural component of healing, when protracted, it can lead toincreased pain and delayed or abnormal healing. In contrast, CaM/eNOS/NOsignaling has been shown to attenuate levels of IL-1β and down-regulateiNOS. As tissue further responds to injury, the CaM/NO/cGMP cascade isactivated in endothelial cells to stimulate angiogenesis, without whichnew tissue growth cannot be sustained. Evidence that non-thermal EMF canmodulate this cascade is provided by several studies. An early studyshowed that the original BGS signal promoted the creation of tubular,vessel-like, structures from endothelial cells in culture in thepresence of growth factors. Another study using the same BGS signalconfirmed a seven-fold increase in endothelial cell tubularization invitro. Quantification of angiogenic proteins demonstrated a five-foldincrease in FGF-2, suggesting that the same BGS signal stimulatesangiogenesis by increasing FGF-2 production. This same study alsoreported increased vascular in-growth more than two-fold when applied toan implanted Matrigel plug in mice, with a concomitant increase inFGF-2, similar to that observed in vitro. The BGS signal significantlyincreased neovascularization and wound repair in normal mice, andparticularly in diabetic mice, through an endogenous increase in FGF-2,which could be eliminated by using a FGF-2 inhibitor.

Similarly, a pulse modulated radio frequency (PRF) signal of the typeused clinically for wound repair was reported to significantlyaccelerate vascular sprouting from an arterial loop transferred from thehind limb to the groin in a rat model. This study was extended toexamine free flap survival on the newly produced vascular bed. Resultsshowed 95% survival of PRF-treated flaps compared to 11% survival in thesham-treated flaps, suggesting a significant clinical application forPRF signals in reconstructive surgery.

In some embodiments, the proposed EMF transduction pathway relevant totissue maintenance, repair and regeneration, begins withvoltage-dependent Ca²⁺ binding to CaM, which is favored when cytosolicCa²⁺ homeostasis is disrupted by chemical and/or physical insults at thecellular level. Ca/CaM binding produces activated CaM that binds to, andactivates, cNOS, which catalyzes the synthesis of the signaling moleculeNO from L-arginine. This pathway is shown in its simplest schematic formin FIG. 77A.

As shown in FIG. 77A, cNOS* represents activated constitutive nitricoxide synthase (cNOS), which catalyzes the production of NO fromL-arginine. The term “sGC*” refers to activated guanylyl cyclase whichcatalyzes cyclic guanosine monophosphate (cGMP) formation when NOsignaling modulates the tissue repair pathway. “AC*” refers to activatedadenylyl cyclase, which catalyzes cyclic adenosine monophosphate (cAMP)when NO signaling modulates differentiation and survival.

According to some embodiments, an EMF signal can be configured toaccelerate cytosolic ion binding to a cytosolic buffer, such as Ca²⁺binding to CaM, because the rate constant for binding, k_(on) isvoltage-dependent and k_(on) is much greater than the rate constant forunbinding, k_(off), imparting rectifier-like properties to ion-bufferbinding, such as Ca²⁺ binding to CaM.

For example, EMF can accelerate the kinetics of Ca²⁺ binding to CaM, thefirst step of a well characterized cascade that responds to chemical orphysical insults. Ca/CaM binding is kinetically asymmetrical, i.e., therate of binding exceeds the rate of dissociation by several orders ofmagnitude (k_(on)>>k_(off)), driving the reaction in the forwarddirection. Ca/CaM binding has been well characterized, with the bindingtime constant reported to be in the range of 10⁻²-10⁻³ sec. In contrast,release of Ca²⁺ from CaM cannot occur until cNOS* has convertedL-arginine to citrulline and NO, which takes the better part of asecond. Subsequent reactions involving NO depend upon the cell/tissuestate. For example, tissue repair requires a temporal sequence ofinflammatory, anti-inflammatory, angiogenic and proliferativecomponents. Endothelial cells orchestrate the production of FGF-2 andVEGF for angiogenesis. For each of these phases, early NO production byendothelial cells, leading to increased cGMP by these, as well as otherNO targets, such as vascular smooth muscle, would be expected to bemodulated by an EMF effect on sGC via Ca/CaM binding. In contrast, nerveor bone regeneration may require other pathways leading todifferentiation during development and growth, and prevention ofapoptosis, as in response to injury or neurodegenerative diseases. Forthese cases, early cyclic adenosine monophosphate (cAMP) formation wouldbe modulated by an EMF effect on sAC via Ca/CaM binding.

The substantial asymmetry of Ca/CaM binding kinetics provides a uniqueopportunity to configure EMF signals that selectively modulate k_(on).In general, if k_(on)>>k_(off), and k_(on) is voltage-dependent,according to the present invention, ion binding could be increased withan exogenous electric field signal having a carrier period or pulseduration that is significantly shorter than the mean lifetime of thebound ion. This applies to the CaM signaling pathway, causing it toexhibit rectifier-like properties, i.e., to yield a net increase in thepopulation of bound Ca²⁺ because the forward (binding) reaction isfavored. The change in surface concentration, ΔΓ, of Ca²⁺ at CaM isequal to the net increase in the number of ions that exit the outerHelmholtz plane, penetrate the water dipole layer at the aqueousinterface of the binding site, and become bound in the inner Helmoltzplane. For the general case of ion binding, evaluation of Ca/CaM bindingimpedance, ZA(s), allows calculation of the efficacy of any givenwaveform in that pathway by evaluating the frequency range over whichthe forward binding reaction can be accelerated. Thus, binding current,IA(t), is proportional to the change in surface charge (bound ionconcentration) via dq(t)/dt, or, in the frequency domain, via sqA(s).IA(s) is, thus, given by:

I _(A)(s)=sq _(A)(s)=sΓ _(o) f(ΔΓ(s))  (1)

where s is the real-valued frequency variable of the Laplace transform.Taking the first term of the Taylor expansion of equation 1 gives:

I _(A)(s)=q _(Ì) sÌ _(o)

Ì(s)  (2)

where qΓ=∂q/∂F, a coefficient representing the dependence of surfacecharge on bound ion concentration. ΔΓ(s) is a function of the appliedvoltage waveform, E(s), and, referring to the reaction scheme in FIG.77, of the change in concentration of eNOS*, defined as ΔΦ(s):

ΔΓ(s)=k _(on)/Γ_(o) s[−ΔΓ(s)+aE(s)+ΔΦ(s)]  (3)

where Γ_(o) is the initial surface concentration of Ca²⁺ (homeostasis),and a=∂F/∂E, representing the voltage dependence of Ca²⁺ binding.Referring to the reaction scheme in FIG. 77, it may also be seen thateNOS* depends only upon Ca²⁺ binding, i.e., ΔΓ(s). Thus:

ΔΦ(s)=υΦ/Φ_(os)[−ΔΦ(s)−ΔΓ(s)]  (4)

where υΦ is the rate constant for Ca/CaM binding to eNOS and Φo is theinitial concentration of eNOS* (homeostasis).

Using equations 2, 3 and 4, and for k_(on)>>υ_(φ), ZA(s) may be written:

$\begin{matrix}{{Z_{A}(s)} = {\frac{E(s)}{I_{A}(s)} = {\frac{1}{q_{\Gamma}a}\lbrack \frac{1 + {\Gamma_{o}{s/k_{on}}}}{\Gamma_{o}s} \rbrack}}} & (5)\end{matrix}$

Equation 5 describes the overall frequency response of the first bindingstep in a multistep ion binding process at an electrified interface,wherein the second step requires that the bound ion remain bound for aperiod of time significantly longer than the initial binding step. Forthis case, the first ion binding step is represented by an equivalentelectrical impedance which is functionally equivalent to that of aseries R_(A)-C_(A) electric circuit, embedded in the overall dielectricproperties of the target. R_(A) is inversely proportional to the bindingrate constant (k_(on)), and C_(A) is directly proportional to bound ionconcentration.

Some embodiments provide that a electromagnetic field, for which pulseduration or carrier period is less than about half of the bound ionlifetime can be configured to maximize current flow into the capacitanceCA, which will increase the voltage, E_(b)(s), where s is the LaPlacefrequency, across CA. E_(b)(s) is a measure of the increase in thesurface concentration of the binding ion in the binding sites of thebuffer, above that which occurs naturally in response to a givenphysiological state. The result is an increase in the rate ofbiochemical signaling in plant, animal and human repair, growth andmaintenance pathways which results in the acceleration of the normalphysiological response to chemical or physical stimuli. The followingequation demonstrates the relation between the configuredelectromagnetic waveform, E(s) and E_(b)(s).

$\begin{matrix}{{E_{b}(s)} = \frac{( {1/{sC}_{A}} ){E(s)}}{( {R_{A}^{2} + ( {1/{sC}_{A}} )^{2}} )^{1/2}}} & (6)\end{matrix}$

Some embodiments also provide that a time-varying electromagnetic fieldfor which pulse duration or carrier period is less than about half ofthe bound ion lifetime of Ca²⁺ binding to CaM will maximize the currentflow into the Ca/CaM binding pathway to accelerate the CaM-dependentsignaling which plants, animals and humans utilize for tissue growth,repair and maintenance. In particular, a time-varying electromagneticfield may be configured to modulate CaM-dependent NO/cGMP signalingwhich accelerates; pain and edema relief, angiogenesis, hard and softtissue repair, repair of ischemic tissue, prevention and repair ofneurodegenerative diseases, nerve repair and regeneration, skeletal andcardiac muscle repair and regeneration, relief of muscle pain, relief ofnerve pain, relief of angina, relief of degenerative joint disease pain,healing of degenerative joint disease, immunological response todisease, including cancer.

Another embodiment according to the present invention is anelectromagnetic signal which accelerates the kinetics of Ca²⁺ binding bymaximizing non-thermal E_(b)(s) at its CaM binding sites, consisting ofa 1-10 msec pulse burst of 27.12 MHz radio frequency sinusoidal waves,repeating between about 1 and about 5 bursts/sec and inducing a peakelectric field between about 1 and about 100 V/m, then coupling theconfigured waveform using a generating device such as ultra lightweightwire coils that are powered by a waveform configuration device such asminiaturized electronic circuitry which is programmed to apply thewaveform at fixed or variable intervals, for example 1 minute every 10minutes, 10 minutes every hour, or any other regimen found to bebeneficial for a prescribed treatment.

In some embodiments, the PEMF signal configuration used may be asinusoidal wave at 27.12 MHz with peak magnetic field B=0.05 G(Earth=0.5 G), burst width, T1=5 msec, and repetition rate T2=2/sec asshown in FIG. 86A. The PEMF signal configuration may also induce a 1-5V/m peak electric field in situ with a duty cycle=2%, without heat orexcitable membrane activity produced. The field may be applied throughan electrical pulse generator to a coil tuned to 27.12 MHz. The burstwidth and repetition rate may be chosen by comparing the voltage inducedacross the Ca²⁺ binding site over a broad frequency range to noisefluctuations over the same range. Effects of burst widths of two 27.12MHz sinusoidal signals at 1 Hz are illustrated in FIG. 86B. As shown inFIG. 10B, high signal-to-noise ratios (SNRs) can be achieved in therelatively low frequency range and at peak magnetic field 0.05 G.

FIG. 78A illustrates a block diagram of an EMF delivery apparatus asdescribed according to some embodiments. As shown in FIG. 78A, theapparatus may have miniaturized circuitry for use with a coilapplicator. In some embodiments, the apparatus may include a CPUMODULATOR, a BATTERY MODULE, a POWER SUPPLY, On/Off switch, and anoutput amplifier, AMP, as illustrated. In further variations, the CPUMODULATOR may be an 8 bit 4 MHz micro-controller; however, othersuitable bit-MHz combination micro-controllers may be used as well. Forexample, in some embodiments, the CPU MODULATOR may be programmed for agiven carrier frequency or pulse duration, such as about 27.12 MHzsinusoidal wave. Moreover, the CPU MODULATOR may be programmed for agiven burst duration, for example about 3 msec. In further variations,the CPU MODULATOR may be programmed to provide a given in situ peakelectric field, for example 20 V/m; or a given treatment time, forexample about 15 minutes; and/or a given treatment regimen, for exampleabout 10 minutes about every hour. The CPU MODULATOR may also beprogrammed to deliver an EMF waveform to the target ion binding pathway.

In further embodiments, the BATTERY MODULE may be rechargeable. In otherembodiments, the BATTERY MODULE has an output voltage of 3.3 V; however,other voltages can be used as is understood in the art. In furthervariations, the BATTERY MODULE supplies DC voltage and current to aPOWER SUPPLY which provides operating power to the CPU MODULATOR and theoutput amplifier AMP.

In some variations, the electromagnetic signal (or a field generatedfrom a electromagnetic signal) is applied inductively to the plantanimal or human target with a COIL applicator, or capacitively withelectrodes in electrochemical contact with the out conductive surface ofthe target structure (not shown). In some variations, the COILapplicator is flexible and circular, but may also be anatomicallyconformable, such as oval or saddle shaped, with a diameter of betweenabout 2 cm to about 50 cm. An electromagnetic treatment, or, if desired,an electromagnetic treatment regimen, can be initiated with the ON/OFFswitch, which may be mechanical or electronic.

Some embodiments combine the signal generation and coil or electrodeapplicator into one portable or disposable unit, such as illustrated inFIG. 78B (which will be described in greater detail below) for the caseof an inductively coupled signal. In some variations, when electricalcoils are used as the applicator, the electrical coils can be poweredwith a time varying magnetic field that induces a time varying electricfield in a target pathway structure according to Faraday's law. Anelectromagnetic field generated by a circuit such as shown in FIG. 78Acan also be applied using electrochemical coupling, wherein electrodesare in direct contact with skin or another outer electrochemicallyconductive boundary of a target pathway structure.

In yet another embodiment, the electromagnetic field generated by thegenerating circuit of FIG. 78A (or FIG. 78B) can also be applied usingelectrostatic coupling wherein an air gap exists between a generatingdevice such as an electrode and a target pathway structure such as amolecule, cell, tissue, and organ of a plant animal or human.Advantageously, the ultra lightweight coils and miniaturized circuitry,according to some embodiments, allow for use with common physicaltherapy treatment modalities and at any location on a plant, animal orhuman for which any therapeutic or prophylactic effect is desired. Anadvantageous result of application of some embodiments described is thata living organism's wellbeing can be maintained and enhanced.

Referring to FIG. 78C, an embodiment according to the present inventionof an induced electric field waveform delivered to a target pathwaystructure is illustrated. As shown in FIG. 78C, burst duration andperiod are represented by T₁ and T₂, respectively. In some embodiments,the signal within the rectangular box designated at T₁ can be,rectangular, sinusoidal, chaotic or random, provided that the waveformduration or carrier period is less than one-half of the target ion boundtime. The peak induced electric field is related to the peak inducedmagnetic field, shown as B in FIG. 78C, via Faraday's Law of Induction.

In further variations, the induced electric field waveform provides aburst of duration between about 1 msec and about 30 msec, containing arepetitive rectangular pulse, a sinusoidal wave or a chaotic or randomwaveform, having, respectively, a period or frequency less than half ofthe bound time of the target ion binding pathway, repeats between about1 and about 10 bursts/sec, and induces a peak electric field of 20 V/mwhich is proportional to a peak applied time varying magnetic field of50 mG according to Faraday's Law of Induction. The induced electricfield illustrated in FIG. 78C can be configured according to embodimentsdescribed to modulate biochemical signaling pathways in plant, animaland human targets, such as those illustrated in FIG. 77A.

In addition to the above, induced time-varying electric fields (e.gPEMF) may be configured to affect neurological tissue including specificcellular/molecular pathways in the CNS tissues allowing these tissues toreact in a physiologically meaningful manner. For example, a waveformmay be configured within a prescribed set of parameters so that aparticular pathway, such as CaM-dependent NO synthesis within theneurological tissue target, is modulated specifically. Both the appliedwaveform and the dosing or treatment regime applied may be configured sothat at least this pathway is targeted specifically and effectively.Furthermore, the stimulation protocol and dosing regimen may beconfigured so that an electromagnetic field applicator device may beportable/wearable, lightweight, require low power, and does notinterfere with medical or body support such as wound dressings,orthopedic and other surgical fixation devices, and surgicalinterventions.

In some embodiments, a method of treating a subject for a neurologicalcondition, injury, or disease includes applying the one or more (or arange of) waveforms that are needed to target the appropriate pathwaysin the target neuronal tissue. This determination may be made throughcalculation of mathematical models such as those described in U.S.Patent Publication No. 2011-0112352 filed Jun. 21, 2010 as U.S. patentapplication Ser. No. 12/819,956 (herein incorporated by reference) todetermine the dosing regimen appropriate for modulating a molecularpathway (e.g. Ca/CaM pathway).

For example, as discussed above, it is believed that pathways involvedin the maintenance and repair of cerebral tissue include the Ca/CaMpathway. To modulate this pathway, in some variations, theelectromagnetic/fields applied are configured to comprise bursts of atleast one of sinusoidal, rectangular, chaotic or random wave shapes;burst duration less than about 100 msec, with frequency content lessthan about 100 MHz at 1 to 100 bursts per second. In other variations,the electromagnetic fields have a 1 to about a 50 msec burst of radiofrequency sinusoidal waves in the range of about 1 to about 100 MHz,incorporating radio frequencies in the industrial, scientific, andmedical band, for example 27.12 MHz, 6.78 MHz, or 40.68 MHz, repeatingbetween about 0.1 to about 10 bursts/sec. In further variations, a PEMFsignal can be applied that consists of a 2 msec burst of 27.12 MHzsinusoidal waves repeating at 2 Hz. In additional embodiments, anapplied PEMF signal can consist of a sinusoidal waveform of 27.12 MHzpulse-modulated with 4 msec bursts having an amplitude of 0.001 G to 0.1G, and repeating at 2 Hz. In additional embodiments, electromagneticfields applied are configured to have a frequency content in a range ofabout 0.01 Hz to about 10,000 MHz having a burst duration from about0.01 to about 100 msec, and having a burst repetition rate from about0.01 to about 1000 bursts/second.

Alternatively, the carrier signal frequency may be below 1 MHz, such as100,000 Hz, 10,000 Hz, 100 Hz or 1 Hz. In such variations, the lowercarrier signal frequency requires a longer burst duration, e.g. 500 msecfor 100 Hz carrier frequency, and a lower amplitude of between about0.001 G and 0.01 G.

Electromagnetic signals can be applied manually or automatically throughapplication devices to provide a range of electromagnetic fields,treatment ranges and doses. For example, PEMF signals can be applied for15 minutes, 30 minutes, 60 minutes, etc. as needed for treatment.Electromagnetic signals can also be applied for repeated durations suchas for 15 minutes every 2 hours. Treatment duration can also spanminutes, days, weeks, etc. For example, PEMF signals can be applied for15 minutes every 2 hours for 9 days. Furthermore, PEMF treatment can beprovided for a therapeutic period of time. As used herein, the termtherapeutic period is not limiting to any specific treatment regimen,but rather describes at least the total treatment period and treatmentperiod per each treatment cycle. For example, a PEMF signal may beapplied for 15 minutes every 2 hours continuously until levels ofintracranial pressure decrease to acceptable levels. The therapeuticperiod would include at least the treatment interval, anyinter-treatment interval, and the total treatment duration.

The electromagnetic applicator devices can also provide a time varyingmagnetic field (for example, peak=0.001 G to 0.1 G, Average=10⁻⁶ G to10⁻³ G) to induce a time varying electric field (for exampleaverage=0.1V/m to 100V/m) in the tissue target. Moreover, each signalburst envelope may be a random function providing a means to accommodatedifferent electromagnetic characteristics of target tissue. Similarly,the number of treatments and the dose regime may vary depending on theprogress of the target location.

In some embodiments, modifying neuronal pathways can result in increasedor decreased cerebral blood flow to a target location. For example,modulating the Ca/CaM pathway can cause vasodilation in the targetcerebral tissue. Vasodilation of cerebral tissue can result in increasedcerebral blood flow which can mitigate inflammation, neuronaldegeneration, and tissue death and promote tissue regrowth, repair, andmaintenance.

In further embodiments, PEMF can be configured to treat a subject havinga metal implant or other foreign object affixed to or penetrating theskull such that the treatment is not affected by the foreign object.Dose regimens such as those described above may still be applied in thepresence of foreign metal objects that may have penetrated the skull(e.g. shrapnel) or been implanted (e.g. skull plate) by carefulpositioning of the applicator coil with respect to the position of themetal in the target, which advantageously allows for treatment ofsubjects with these conditions.

As is understood by one of ordinary skill in the art, the termsneurological condition, disease, injury etc. as used herein are notintended to be limited to any particular condition or injury described.A neurological injury can mean at least an injury that results frommechanical damage arising from an initial insult or trauma event and/orany secondary injury from secondary physiological responses. In someembodiments, the methods and devices contemplated may be configured totreat patients for whom the trauma event is initiated by medicalpersonnel as part of another treatment. For example, in the case of acraniotomy to remove brain tumors or lesions, the neurological injurywould include the surgical incision(s) into brain tissue and subsequentsecondary injury from resulting inflammation or swelling that developsafter the initial insult. Similarly, neurological conditions or diseasescan mean at least, and non-exhaustively, degenerative disorders such asAlzheimer's or neurological, functional, or behavioral impairment(s)resulting from injury. For example, secondary physiological responsessuch as inflammation can damage healthy brain tissue which can result inimpairment of a cognitive or behavioral function associated with thatpart of the brain.

FIG. 77B is a flow diagram of a method for treating a subject with aneurological condition, disease, or injury. In some variations, beforebeginning the treatment, one or more (or a range of) waveforms may bedetermined that target the appropriate pathway for the target tissue. Insuch embodiments, once this determination is made, electromagneticfields are applied to the target location.

In further embodiments, the treatment waveform or PEMF signal may bedetermined by configuring the PEMF waveform to target a rhythm patternof a physiological system or process. For example, a PEMF signal may beconfigured to modulate brain rhythms to effect relaxation or alertnessdepending on the needed physiological response. As is understood in theart, physiological systems like the CNS and the peripheral nervoussystem (PNS), in particular, the brain or heart emit electrical activitythat can be measured and recorded by, for example,electroencephalography (EEG) or electrocardiography (EKG). Duringparticular activities, such as sleep/rest or problem solving, the brainemits electrical/rhythmic activity (e.g. circadian rhythms) in certainfrequency bands associated with the activity (e.g. theta, alpha, beta,etc.)

A PEMF waveform can be configured to a specific rhythm of a targetlocation by providing a signal with the frequency, amplitude, burstduration, etc. associated with a particular activity of that targetlocation. For example, for treatment of a neurological condition such asAlzheimer's, a PEMF waveform can be brought in close proximity to aregion of the brain associated with problem solving. In such cases, thePEMF waveform provided to the patient can be configured to the rhythmfrequency/band that is generally measured when normal problem solvingskills are employed. The PEMF waveform may then be used to stimulate thetarget region while the patient is engaged in a problem solvingactivity. This treatment may help the patient regain or improve problemsolving skills where the target region has exhibited diminished abilityto emit normal electrical activity.

In further embodiments, the PEMF waveform may be configured to modulaterhythms associated with a physiological response that arises from aneurological injury. For example, as can be appreciated, neurologicaldamage such as traumatic brain injury results in both secondaryphysiological responses in the CNS as well as responses in peripheralsystems. With brain trauma, a patient's ability to regulate and maintainperiphery systems such as the cardiac and pulmonary systems may beindirectly compromised. As such, some embodiments contemplated providefor PEMF configurations that treat a neurological injury by targetingnon-neurological systems affected by the injury. In some embodiments,the PEMF waveforms are configured to modulate the rhythms or electricalactivity of one or more non-neurological system(s).

In further embodiments, the PEMF waveform may be configured to modulatesleep patterns. In particular, PEMF configurations may increase theduration of slow-wave (Delta) sleep in each sleep cycle which may allowthe injured person to maximize the production of human growth hormone,which, in turn, may increase healing for any injury, including CNS andPNS injuries, and provides a prophylactic response to protect fromfurther injury.

As described in FIG. 77B, a method of treating a subject with aneurological injury or condition may include the step of placing thetissue to be treated (e.g. near one or more CNS regions) in contact, orin proximity to, a PEMF device 77101. Any appropriate PEMF device may beused. In general, the device may include an applicator (e.g. inductorapplicator) which may be placed adjacent to or in contact with thetarget location/tissue. The device may also contain a signalconditioner/processor for forming the appropriate waveform toselectively and specifically modulate a pathway (e.g. Ca/CaM pathway).In further embodiments, the device may include a timing element (e.g.circuit) for controlling the timing automatically after the start of thetreatment.

In the example shown in FIG. 77B, once treatment begins 77103, thedevice, in some variations, applies an envelope of high-frequencywaveforms at low amplitude (e.g. less than 50 milliGauss, less than 100milliGaus, less than 200 milliGauss, etc.) 77105. This envelope ofhigh-frequency pulses is then repeated at a particular frequency afteran appropriate delay. This series of bursts can be repeated for a firsttreatment time (e.g. 5 minutes, 15 minutes, 20 minutes, 30 minutes,etc.) and then followed by a delay during which the treatment is “off”77107. This waiting interval (inter-treatment interval) may last forminutes or hours (15 minutes, 2 hours, 4 hours, 8 hours, 12 hours, etc.)and then the treatment interval may be repeated again until thetreatment regime is complete 77109.

In some variations, the treatment device is pre-programmed (orconfigured to receive pre-programming) to execute the entire treatmentregime (including multiple on-periods and/or intra-treatment intervals)punctuated by predetermined off-periods (inter-treatment intervals) whenno treatment is applied. In further variations, the device ispre-programmed to emit a PEMF signal at 27.12 MHz at 2 msec burstsrepeating at 2 bursts/sec. In other embodiments, the device ispre-programmed to emit a PEMF signal at 27.12 MHz (at about amplitude250-400 mV/cm) pulsed in 4 msec bursts at 2 Hz.

As discussed, the selection of a treatment regime may be determined bythe particular neurological injury or condition etc. at issue. In thecase of treating secondary physiological responses from TBI, thetreatment parameters may be selected to target any number or combinationof physiological responses. For example, some embodiments contemplatedprovide for devices and methods for reducing intracranial pressure.Oftentimes a trauma event such as brain surgery will induce cerebraledema, the extra- and intracellular accumulation of fluid resulting fromchanges in vascular endothelium causing vasodilation and leakage as wellas surges of extracellular fluid into cells after disturbances inglutamate release and calcium and sodium ion influx. This is potentiallyfatal as increased intracranial pressure decreases cerebral perfusionpressure and interrupts cerebral blood flow to brain tissue, which cancause ischemia and neuronal death.

To manage intracranial pressure, some embodiments provide a method ofreducing intracranial pressure by applying a PEMF signal in closeproximity to a target location. Such treatment parameters may includeany of those discussed, which are found suitable for the needs of thepatient. Moreover, in some embodiments, the selected PEMF signal can beapplied continuously to the target area until an acceptable intracranialpressure level is reached. An acceptable intracranial pressure level canbe patient-specific depending on the circumstances; however, generallynormal intracranial pressure ranges from about 5 mmHg to about 15 mmHg.Additionally, intracranial pressure above about 20 mmHg is generallyconsidered harmful. As such, PEMF treatment may be initiated onceintracranial pressure is above an acceptable level.

Alternatively, PEMF treatment may be discontinued once acceptable levelsare attained. In some embodiments, the PEMF treatment can be applied asshown in FIG. 77B with inter-treatment intervals. For example, a PEMFsignal of 27.12 MHz pulsed in 4 msec bursts at 2 Hz may be applied for15 minutes every 2 hours for 9 days. In other embodiments, the PEMFsignal may be applied continuously without an inter-treatment intervaluntil an acceptable level of intracranial pressure is reached. Infurther embodiments, the PEMF therapy includes monitoring a neurologicalfactor such as intracranial pressure of the subject such that PEMFtreatment can be initiated or discontinued depending on the levels ofintracranial pressure.

In some embodiments, the patient may experience intracranial pressurebelow about 20 mmHg; however, due to lower cerebral perfusion pressure,PEMF therapy may be initiated to mitigate conditions such as ischemia.In further embodiments, the PEMF therapy may be preventative and appliedto maintain the subject's pressure levels within a selected range thatmay or may not be within the normal pressure ranges described above.Additionally, a PEMF device may be pre-programmed with a controller orprocessor that monitors and adjusts PEMF treatment based on the levelsof intracranial pressure. A PEMF device may be configured to communicatewith a sensor or other data gathering devices/components that provideinformation regarding intracranial pressure or other neurologicalfactors.

In addition to intracranial pressure, additional embodiments provide forPEMF methods and devices for treating inflammation resulting as asecondary physiological response to neurological injury (e.g. TBI)Inflammation is a natural and protective systemic physiological responseto invading pathogens to preserve tissue viability and function.However, if this process remains unchecked, it can lead to secondarytissue damage in the CNS. In the case of brain injury, inflammation canrestrict cerebral blood flow and cause damage or death to healthy braintissue. Although the complex process involved in inflammation is notcompletely known, it is understood that following injury, microglia andastrocytes will activate and migrate to the injury site. Once activated,these cells will secrete destructive cytokines (e.g. IL-la, IL-1β andTNF-α) as well as other inflammatory molecules such as chemokines, whichcan attract additional immune-mediators. Some of these immune-mediatorscan penetrate the blood-brain barrier and further add to an inflammatoryresponse. Although microglia, cytokines, chemokines, and otherinflammatory promoters are required to some extent to remove invadingpathogens, protracted and unremitting inflammation can cause long termdamage. As such, some embodiments provide for PEMF treatments anddevices to alter the levels of inflammatory factors present in a targetlocation.

Because increased levels of cytokines such as IL-1β have been correlatedwith high intracranial pressure, inflammation, and breakdown of theblood-brain barrier, some embodiments provide for a PEMF treatment thatcan reduce or mitigate the levels of cytokines in order to preventsecondary injury to target brain tissue. In such embodiments, a PEMFapplicator device such as the one described in FIG. 78B is placed inclose proximity to a target tissue location (e.g. brain area). The PEMFapplicator device is then activated and generates a PEMF signalconfigured to reduce the levels of cytokines in the target location. Insome embodiments, a PEMF signal of 27.12 MHz pulsed in 4 msec bursts at2 Hz for about 5 to about 15 minutes every 20 minutes to reduce thequantities of IL-1β present in target location. The PEMF signal may beapplied for a selected amount of time before pausing for aninter-treatment interval (see FIG. 77B) and then repeated for a totaltreatment time. In further embodiments, the PEMF treatment may beapplied continuously until acceptable levels of cytokines orinflammation are reached. The PEMF treatment may also be appliedcontinuously or intermittently in response to direct data regardinglevel of cytokines or inflammation or indirect data such as levels ofcerebral blood flow.

In other embodiments, the PEMF treatment may be directed toward alteringthe levels of microglia or astrocytes present in the target location. Asdiscussed, once activated, microglia not only produce cytokines but alsoremove damaged or dead tissue and infectious agents. In other words,microglia are dually neuroprotective and neurotoxic. As such, reducingor increasing the levels of microglia at different stages following aneurological injury or condition may modulate the helpful and harmfuleffects of microglia present in the target location. For example, in theimmediate period following injury, an increase in activated microgliamay help to clear and collect pathogens and debris from cellular ortissue damage. By doing so, an increased level of microglia can reducethe chances of infection and prevent inflammation before it begins.Moreover, increased activity of microglia may enhance the repair ofaxons. Alternatively, at a later stage post-injury, reducing the numberof activated microglia can reduce inflammation by preventing microgliafrom producing pro-inflammatory factors such as cytokines andchemokines. As the suitability of increasing or decreasing microglialevels in a target area are dependent on the type of injury/conditionand the patient's needs, flexibility will be needed to modify PEMFtreatment as needed. In some embodiments, the PEMF device/treatment canbe pre-programmed to alter treatment as needed according to monitoredconditions such as the levels of inflammation, levels of microglia, ortime period after injury. In other variations, the PEMF treatment can bemanually modified as needed. In further variations, the PEMF treatmentmay appear to first decrease microglial activity, but the apparentdecrease in microglia may be transitory and microglial activity mayactually be increased/accelerated over the course of treatment. As shownin FIGS. 87 and 88 (and further described in detail in Examples 7 and8), PEMF treatment can effect an increase or decrease in microglialactivity.

Further embodiments provide for treatments and devices for preventingneuronal death. Injuries caused by both contusive trauma and by invadingforeign objects (e.g. penetrating injury) will kill neurons, which canbe responsible for lasting behavioral deficits as well as limbic andcognitive disabilities. Some PEMF treatments contemplated provide fortherapies that increase neuronal survival. For example, PEMF signals canbe applied to a target location with damaged neuronal cells. The PEMFsignals may increase the level of activated microglia present at thesite, for example, which can help remove pathogens that could causeinfection to already damaged neuronal cells. Moreover, reductions intissue swelling and inflammation also indirectly increase neuronalsurvival, as these processes can both initiate and exacerbate acute andchronic neurodegeneration. Treatment parameters may be selectedaccording any of the described regimes as needed for treatment.

In treating neurological conditions and injuries, a primary concern isretaining or recovering cognitive, motor, limbic, and behavioralfunctions. Tissue damage and death, especially in the brain, canirreversibly affect the ability of patients to function normally after atraumatic event. Some embodiments provide for treatments and devices toimprove cognitive, motor, behavioral etc. function after a neurologicalinjury/condition. Some variations provide for short term and long termPEMF treatment where ongoing assessment of the patient's progress isrecorded to determine whether treatment should be continued or modified.

As can be appreciated, PEMF signals can be configured to treat one ormore of the conditions described. For example, a PEMF treatment may beused to reduce intracranial pressure and inflammation in a patient inneed thereof.

FIG. 78B illustrates an embodiment of an apparatus 78200 that may beused. The apparatus is constructed to be self-contained, lightweight,and portable. A circuit control/signal generator 78201 may be heldwithin a (optionally wearable) housing and connected to a generatingmember such as an electrical coil 78202. In some embodiments, thecircuit control/signal generator 78201 is constructed in a manner thatgiven a target pathway within a target tissue, it is possible to choosewaveform parameters that satisfy a frequency response of the targetpathway within the target tissue. For some embodiments, circuitcontrol/signal generator 78201 applies mathematical models or results ofsuch models that approximate the kinetics of ion binding in biochemicalpathways. Waveforms configured by the circuit control/signal generator78201 are directed to a generating member 78202. In some variations, thegenerating member 78202 comprises electrical coils that are pliable andcomfortable. In further embodiments, the generating member 78202 is madefrom one or more turns of electrically conducting wire in a generallycircular or oval shape, any other suitable shape. In further variations,the electrical coil is a circular wire applicator with a diameter thatallows encircling of a subject's cranium. In some embodiments, thediameter is between approximately 6-8 inches. In general, the size ofthe coil may be fixed or adjustable and the circuit control/signalgenerator may be matched to the material and the size of the applicatorto provide the desired treatment.

The apparatus 78200 may deliver a pulsing magnetic field that can beused to provide treatment of a neurological condition or injury. In someembodiments, the device 78200 may apply a pulsing magnetic field for aprescribed time and can automatically repeat applying the pulsingmagnetic field for as many applications as are needed in a given timeperiod, e.g. 6-12 times a day. The device 78200 can be configured toapply pulsing magnetic fields for any time repetition sequence. Withoutbeing bound to any theory, it is believed that when electrical coils areused as a generating member 78202, the electrical coils can be poweredwith a time varying magnetic field that induces a time varying electricfield in a target tissue location.

In other embodiments, an electromagnetic field generated by thegenerating member 78202 can be applied using electrochemical coupling,wherein electrodes are in direct contact with skin or another outerelectrically conductive boundary of the target tissue (e.g. skull orscalp). In other variations, the electromagnetic field generated by thegenerating member 78202 can also be applied using electrostatic couplingwherein an air gap exists between a generating member 78202 such as anelectrode and the target tissue. In further examples, a signal generatorand battery is housed in the miniature circuit control/signal generator78201 and the miniature circuit control/signal generator 78201 maycontain an on/off switch and light indicator. In further embodiments,the activation and control of the treatment device may be done viaremote control such as by way of a fob that may be programmed tointeract with a specific individual device. In other variations, thetreatment device further includes a history feature that records thetreatment parameters carried out by the device such that the informationis recorded in the device itself and/or can be transmitted to anotherdevice such as computer, smart phone, printer, or other medicalequipment/device.

In other variations, the treatment device 78200 has adjustabledimensions to accommodate fit to a variety of patient head sizes. Forexample, the generating member 78202 may comprise modular componentswhich can be added or removed by mated attaching members. Alternatively,the treatment device 200 may contain a detachable generating member(e.g. detachable circular coil or other configurations) that can beremoved and replaced with configurations that are better suited for theparticular patient's needs. A circular coil generating member 78202 maybe removed and replaced with an elongate generating member such thatPEMF treatment can be applied where other medical equipment may obstructaccess by a circular generating member 78202. In other variations, thegenerating member may be made from Litz wire that allows the generatingmember to flex and fold to accommodate different target areas or sizes.

In other embodiments, the diameter of a circular generating member maybe selected based on the desired treatment regimen. In some variations,the depth of penetration for the electromagnetic field increases withincreased diameter. In such embodiments, a larger diameter will providea field with a greater field volume allowing for greater penetration inthe target location. Accordingly, by modifying the diameter or size ofthe generating member, the depth of the treatment field can be adjustedas needed. Greater depth of penetration may be advantageous where theinjured target region is below the surface of the target location.Alternatively, where a greater depth of penetration is not needed,generating members of smaller size may be more appropriate where surfaceapplication is desired. For example, for treatment of a large surfacearea, an array of smaller sized generating members can be used to covera large area without deep penetration beyond the surface.

In further embodiments, the inductive device illustrated in FIG. 78B isflexible, portable and, if desired, disposable; and can be used alone orincorporated into an anatomical positioning device such as a dressing,bandage, compression bandage, compression dressing; knee, elbow, lowerback, shoulder, foot, and other body portion wrap and support; garments,footwear, gloves, and fashion accessories; mattress pads, seat cushions,furniture, beds; in seats or beds within cars, motorcycles, bicycles,buses, trains, planes, boats and ships.

In some embodiments, the devices may include a sensor configured tomonitor a patient's condition for changes. For example, a device mayinclude a sensor that collects data on the patient's intracranialpressure. Based on the amount of intracranial pressure, the device mayautomatically turn on for treatment once threshold pressure levels arereached. Similarly, the device may turn off automatically if pressurelevels return to normal. Additionally, a device providing treatment maymodify and adjust treatment parameters based on the feedback fromsensors. For example, a device may change treatment parameters if thesensor registers an increase in intracranial pressure. Moreover, in somevariations, medical staff may be notified of changes to treatmentparameters where the delivery device can communicate with another devicesuch as computer, smart phone, printer, or other medicalequipment/device.

Example 1

An EMF signal, configured according to an embodiment of the presentinvention to modulate CaM-dependent signaling, consisting of a 27.12 MHzcarrier, pulse-modulated with a 3 msec burst repeating at 2 Hz and apeak amplitude of 0.05 G, was applied for 30 minutes to the MN9Ddopaminergic neuronal cell line and increased NO production byseveral-fold in a serum depletion paradigm and produced a 45% increasein cGMP. The EMF effects on NO and cGMP were inhibited by the CaMantagonist N-(6-Aminohexyl)-5-chloro-1-naphthalenesulfonamidehydrochloride (W-7), indicating the EMF signal acted in this neuronalculture according to the transduction mechanism illustrated in FIG. 77A.These results are summarized in FIG. 79A.

The effect of the same EMF signal on cAMP production in MN9D cells wasalso studied. MN9D cells in serum free medium were removed from theincubator (repeatable temperature stress injury to transiently increaseintracellular Ca²⁺) and exposed to EMF for 15 min. cAMP was evaluated incell lysates by ELISA. Results demonstrate that an EMF signal,configured according to an embodiment of the present invention,increased cAMP production by several-fold. Notably, the c-NOS inhibitorL-NAME abolished the PEMF effect on cAMP. The results, summarized inFIG. 79B, indicate EMF signals, configured according to an embodiment ofthe present invention, affect neuronal differentiation and survival.

Example 2

In this example, a highly reproducible thermal myocardial injury wascreated in the region of the distal aspect of the Left AnteriorDescending Artery at the base of the heart of adult male Sprague Dawleyrats. The EMF waveform, configured as an embodiment of the presentinvention, was a 2 msec burst of 27.12 MHz sinusoidal waves repeating at2 bursts/sec delivering 0.05 G at the tissue target. Five freely roaminganimals in a standard rat plastic cage, with all metal portions removed,were placed within a single turn 14×21 inch coil. Exposure was 30 mintwice daily for three weeks. Sham animals were identically exposed, butreceived no EMF signal.

Upon sacrifice, myocardial tissue specimens were stained with CD-31 toevaluate the presence of newly forming blood vessels and capillaries inperi-ischemic tissue. Results at 21 days showed that number of vesselsand capillaries in peri-ischemic myocardial tissue was increased byapproximately 100% (p<0.001) in EMF vs sham exposed animals. That an EMFsignal, configured as an embodiment of the present invention, modulatedCaM-dependent NO release, as illustrated in FIG. 77A, was verified byfeeding animals L-NAME, a cNOS inhibitor, in their drinking water for 7days. EMF, configured as an embodiment of the present invention,accelerated angiogenesis at 7 days by 60%. The EMF effect was abolishedby L-NAME, as illustrated in FIG. 80.

Example 3

In this example, inflammation was induced in the left hind paw of HarlanSprague-Dawley rats (200-340 g) by injection of 100 μL of a 3.5 mg/mLsterile phosphate buffered saline-based carrageenan solution into thefootpad using a 30 gauge tuberculin syringe. The carrageenan dose wascarefully calibrated to produce a mild, controllable form ofinflammation that could be evaluated for rate of onset. Edema wasdetermined using a plethysmometer volume displacement transducer system(Stoelting Company, Wood Dale, Ill.). Edema was measured pre-carrageenaninjection and at 1, 4 and 8 hours post-injection. Rats were exposed toeither the PEMF signal or a control, untreated experimental coilconfiguration for 15 min. EMF exposures were at 0.25, 2, 4 and 8 hourspost-injection. The signal consisted of a 2 msec burst of 27.12 MHzsinusoidal waves repeating at 2 bursts/sec, and inducing 20 V/m electricfield at a target diameter of 2 cm. This PEMF signal was configured,according to an embodiment of the present invention to accelerate Ca²⁺binding in a CaM-dependent signaling pathway. Data were analyzed withSigmaStat 3.0 software (SPSS, Chicago, Ill.) using Student's unpairedt-test and one way ANOVA, as appropriate. Differences were also comparedusing the Mann-Whitney test for two independent groups. Significance wasaccepted at P≦0.05.

The results showed mean edema volume in the sham treated animals was33±7% greater at 1 hour post-injection (P=0.037), 41±8% greater at 4hours (P=0.005), and 47±9% greater at 8 hours (P=0.009) than edemavolume in the PEMF treated animals at these time points. These results,summarized in FIG. 81, demonstrate that a PEMF signal, configured as anembodiment of the present invention, accelerates Ca²⁺ binding to CaM inthe NO signaling cascade that regulates lymphatic evacuation of edemafrom inflammation.

Example 4

In this example, groups of rats were subjected to invasive and contusivetraumatic brain injury and treated with an EMF signal configured as anembodiment of the present invention consisting of a 27.12 MHz carrier,pulse-modulated with a 3 msec burst repeating at 2 Hz and a peakamplitude of 0.05 G.

Adult male Sprague Dawley rats (350-400 g) were housed in aclimate-controlled animal facility with two rats per cage. Food andwater were provided ad libitum in a 12-hour light/dark cycle. Animalswere maintained, operated on, treated, and euthanized in accordance withfederal, state, and IACUC guidelines at the Montefiore Medical Center.

Closed Skull Contusion Injury: Twenty rats (10/group) were subjected toa moderate closed-head injury under anesthesia using the Marmarouimpact-acceleration model, with the following modifications. Briefly,rats were anesthetized with ketamine/medetomidine (0.75 mg/0.5 mg/kg,i.p.). After depilation and disinfection, the calvarium was exposed bycreating a 1 cm vertical, midline incision through the scalp anddisplacing the periosteum. To diffuse the impact force and reduceincidence of skull fracture, a metal washer (10 mm diameter, 2 mmthickness) was affixed directly to the skull with epoxy cement midwaybetween the lambda and bregma. Rats were secured directly underneath theweight-drop device on foam bedding (Foam to Size; Ashland, Va.; springconstant=4.0). A diffuse closed-head injury was produced by dropping a258.7-gram weight in a plexiglass tube from specified heights up to 2meters, creating forces of impact from 1 to 4 Newtons (4.46N). Afterimpact, the disk was removed from the skull and the periosteum and scalpwere approximated with discontinuous nylon sutures. Anesthesia wasreversed and animals were either treated with PEMF signals or placed insimilar containers in the absence of signals.

The Marmarou weight-drop model was selected for this study partlybecause it has been found that the levels of 1L-1β closely correlate tothe force of the injury in the Marmarou weight-drop model. For example,as shown in FIG. 84, in a previous study, rats were subjected to TBIaccording to the Marmarou weight-drop model by varying the height fromwhich a 257 g weight was dropped. After six hours, levels of 1L-1β werequantified in brain tissue by ELISA. Points shown on FIG. 84 representmean values for 3 rats +/−SEM. Data at 0 force was determined from ratsreceiving sham surgery.

Penetrating Brain Injury: Sixty rats were subjected to bilateral stabinjuries to the striatum. Rats were anesthetized withketamine/medetomidine (0.75 mg/0.5 mg/kg, i.p.) and secured on astereotaxic frame (David Kopf) with the tooth bar at 3.3 mm below theinteraural line. After depilation and disinfection, the calvarium wasexposed, as described above, and the separated tissue was secured withhemostats. Two 1 mm burr holes were created by a trephine drill abovethe striatum at stereotactic coordinates 0.5 mm anterior to and 2.5 mmlateral to Bregma. A 23S gauge blunt-end needle from a Hamilton syringewas inserted 5.2 mm below the dura into each striatum and removed overtwo minutes. After lesioning, burr holes were sealed with bone wax andthe incision site was closed with 4-0, non-absorbable nylon sutures.Rats were reversed from anesthesia with 1 mg/kg medetomidine and placedin containers for PEMF treatment.

PEMF treatment: Animals were exposed to PEMF generated by a sinusoidal27.12 MHz radiofrequency signal pulse-modulated with 3 millisecondbursts with 0.05 G amplitude, and repeating at 2 Hz beginningimmediately after surgery from a coil positioned around a plasticshoebox with a ventilated lid and connected to a PEMF signal generatorwhich automatically provided a signal regimen consisting of signal onfor 5 min in every 20 minute time segment for 6 hours. For treatmentslonger than 6 hours, metal cage inserts were removed, food and hydrogelpacks were placed in the cages, and plastic outer cage tops with filterswere placed in a larger container equipped with a metal coil around itsperimeter on a plastic cart in the animal care facility to avoid signaldistortion from surrounding metal.

PEMF signals were delivered externally from a signal generator attachedby a wire to the coil. Treatment was administered for 5 minutes every 20minutes and rats were allowed to move freely in their cages during thistime. Identical procedures were followed for the control group, i.e.rats were placed in identical containers in the same room and were fedand handled in an identical manner to rats receiving treatment.Immediately before euthanasia, rats were re-anesthetized and CSF wascollected, and after euthanasia, brains were harvested and immediatelyeither fixed in 4% paraformaledhyde or frozen at −80° C. until analysis.

CSF Collection: CSF was obtained utilizing a modification of the Nirogitechnique (REF). Briefly, a standard 23 G Vacutainer® Push Button BloodCollection Syringe with 12″ (what is the diameter of the hole) tubing(BD) was connected to a 1 cc insulin syringe. Anesthetized rats werepositioned on a stereotaxic frame with the tooth bar set to angle the athead 45° in a downward direction. The needle was inserted in an uprightposition into the medial portion of the cisterna magna until CSF wasreleased into the tubing. Fluid was collected until blood was visibleand tubing was clamped with a hemostat to separate clear andblood-tainted CSF. Samples of clear CSF were released into microfugetubes and cellular material was pelleted by centrifugation (speed andtime of centrifuge). Cleared samples were immediately frozen at −80° C.

Tissue Processing: For the weight-drop injury, whole brain hemispheresminus cerebella were frozen. For the penetrating injury, a 5-mm cylinderof brain tissue from the left hemisphere surrounding the stab injurywere removed and frozen. The right hemisphere was fixed by immersion in4% paraformaldehyde for 2 days and stored in 30% sucrose with 0.05%sodium azide.

IL-1β Analysis: Frozen specimens were processed by homogenization usinga polytron (model, Manufacturer) in lysis buffer containingtris-buffered saline and centrifuged at 16,000 g for 10 minutes at 4° C.to pellet particulate matter. Supernatants were frozen at −80° C. andtriton X-100 was added to a final concentration of 0.1%. IL-1β levelswere quantified using a rat IL-1β ELISA duo set (R&D Systems) followingmanufacturer's recommendations. Results were normalized for proteincontent determined with the Biorad protein assay.

OX-42 immunohistochemistry: Sixty-micron tissue sections through thestriatum were generated using a tissue slicer (Ted Pella) and storeduntil use in phosphate-buffered saline (150 mM NaCl, 10 mM sodiumphosphate, pH 7.4; PBS) with 0.05% sodium azide. For immunostaining,sections belonging to a series consisting of every 2nd-3rd section werepermeabilized in 0.25% Trition X-100 in PBS with 3% hydrogen peroxide toextinguish endogenous peroxidase activity. Non-specific binding wasblocked with 3% horse serum in PBS with 0.25% Triton X-100 (“block”).Sections were incubated in mouse OX-42 antibody (Serotec; 1:1000) inblock overnight at 37° C. After washing in PBS, sections were incubatedfor 2 hours at 37° C. with biotin anti-mouse in block (Amersham; 1:600).After subsequent washing and 1.5 hours incubation in peroxidase ABC(Vector Laboratories; 1:500), sections were washed and staining wasvisualized with DAB (Vector). After washing, sections were dehydrated,coverslipped, and photographed at 20× magnification.

Evaluation of microglial activation: OX-42 staining was quantified in aseries of every 2-3 sections representing 0.7 mm through the center ofthe lesion for animals subjected to a penetrating lesion. Initially, thedegree of microglial staining in all animals was scored by 4individuals, where scores (0-5) represented staining intensity and thearea occupied by stained cells. Tissue sections from groups that wereexposed to PEMF (null) signals for 24 and 48 hours after injury wereanalyzed further using Image J software. Immuno-stained regions wereoutlined and areas were measured using a calibrated length. Theintensity of immunostaining was quantified by densitometry, defined asan integrated density, calibrated against selected areas from stainedtissue sections that represented the entire spectrum of staining.

Statistical Analysis: Data for each group was compared and analyzed forsignificant differences by Student's t-test and by analysis of variance(ANOVA) followed by Fischer's PLSD test, when more than two groups werecompared. Differences between groups generating p-values equal to orless than 0.05 were considered statistically significant.

PEMF treatment reduced levels of IL-1β after contusive TBI. CSF andbrain tissue were collected from injured animals in PEMF and controlgroups as well as from sham and intact animals after 6 hours, when peakIL-1β levels were expected using this injury model. Results demonstratethat intact animals had the lowest levels of IL-1β (29±4 pg/mg protein),which increased 34% in the sham group to 39±7 pg/mg protein. Afterinjury, mean levels of IL-1β in the group that did not receive PEMFtreatment was 55±3 pg/mg protein, which was not significantly differentfrom levels of this inflammatory cytokine in animals that received shamsurgery. Mean levels of IL-1β in animals that received PEMF treatmentwere 50±4 pg/mg protein, indicating that there were no significanteffects on levels of this peptide in brain homogenates.

Results also demonstrated that levels of IL-1β in CSF changeddramatically in response to both injury and PEMF treatment. Mean levelsof this cytokine in CSF from intact animals was 19±7 pg/mL CSF,increasing to 25±21 pg/mL in the sham group (31%). Levels in animalsreceiving a contusive 3N injury rose to 252±91 pg/mL, a 10-fold increaseover the sham group. Moreover, animals receiving PEMF treatmentdemonstrated significantly lower concentrations (44±25 pg/mL), or levelsthat were 83% lower than those of animals receiving the injury, and lessthan twice the mean concentration of IL-1β in animals receiving shamsurgery.

Results for PEMF reduced levels of IL-1β after penetrating brain injury:Results illustrating the time course of IL-1β expression demonstratesimilarly low levels of IL-1β in brain homogenates from intact and shamanimals; 24±5 and 24±6 pg/mg protein, respectively. In addition, twoanimals from the sham group were treated with PEMF signals for 6 hoursbefore they were euthanized. Animals in this group demonstrated meanIL-1β levels of 15.4 and 16.6 pg/mg protein for PEMF and sham animalsrespectively, but the number of animals in this group was too low tocompare with either intact or sham rats (n=5 and n=2, respectively). At3.5 hours after injury, IL-10 levels increased approximately 2-fold andattained their highest levels of any time point measured at 6 hoursafter injury in PEMF treated and control groups at 93±15 and 99±11 pg/mgprotein, approximately 4 times basal levels. Importantly, at 17 hoursafter injury, levels of IL-1β were significantly lower in the PEMF group(42±5 pg/mg protein) than those of the control group (61±5 pg/mgprotein; p<0.04). Control levels decreased and values at later timepoints were similar in both groups up to 9 days after injury.

In CSF, levels of IL-1β followed a more protracted time course. Inintact naive animals, basal levels of IL-1β were 32±32 pg/mL CSF,demonstrating wide and average levels in the sham group were 56±51 and39±10 pg/mL (control and PEMF-treated, respectively). Levels stayedfairly low at 6 hours after injury, but rose approximately 5-fold toreach a maximum of 224±23 pg/mL at 17 hours after injury, a 7 to 8-foldincrease over basal levels, and to a similar degree as IL-1β levels inCSF from animals receiving the closed-skull contusion. In contrast,animals that received PEMF treatment did not exhibit significantincreases in IL-1β, which was maintained at approximately basal levels(23±18 pg/mL CSF), or ten-fold lower than rats that received an injuryand were not exposed to PEMF signals. Concentrations of IL-1β remainedhigh in the injured control group at 24 hours (122±56 pg/mL), anddecreased to baseline levels at 4 to 9 days after injury (31-45 pg/mL),which persisted throughout this period. IL-1β concentrations were lowestin both groups at 9 days (0-2 pg/mL). Taken together, resultsdemonstrate that PEMF treatment suppressed IL-1β levels in CSFthroughout a 9-day period after penetrating brain injury.

PEMF treatment increased OX-42 expression after penetrating injury. TheCNS responds to focal penetrating injuries by mounting a localinflammatory response. Using the penetrating injury TBI model, theeffects of PEMF treatment on microglial activation were examined.Animals received bilateral penetrating injuries and were assigned toPEMF or null treatment groups, where they received continuous treatmentuntil sacrifice at 3.5 hours to 9 days after lesioning. Resultsdemonstrate that OX-42 staining was absent in the area of the lesion at3.5 and 6 hours. Beginning at 17 hours after injury, OX-42immunoreactivity was detected increasing in intensity and size over 5days. At 9 days, the last time point, staining was most intense andappeared more focal, encompassing the lesion itself a compactedsurrounding area with a well-defined perimeter. Initially, the extent ofstaining was analyzed in a semi-quantitative by rating the intensity andarea of staining on a scale of 1 to 5 in 0.25 increments by four blindedobservers. The overall degree of OX-42 expression, a combination ofstaining intensity and the area of staining, increased over the 9 daysof the experiment.

Significantly, PEMF signals increased the intensity of OX-42 staining at24 and 48 hours after injury. This increase was transient, as valueswere higher, but similar to control levels at both 5 and 9 days afterinjury. The area occupied by OX-42+ cells at 9 days was smaller than at5 days, indicating that microglia had arrived at their destination.Image analysis was employed to confirm our observations. Areas (mm2) andmean gray values (average value of pixels over the area in which OX-42staining was found) were measured on Image J for groups of animalsreceiving PEMF (null) signals for 24 and 48 hours. Interestingly, thearea of OX-42 staining at 24 hours after injury significantly decreasedin the PEMF-treated animals compared to controls, but in contrast, themean intensity of OX-42 immunoreactivity was significantly higher,suggesting that PEMF signals accelerated microglial activation andmigration. The intensity of OX-42 immunoreactivity increased in bothgroups at 48 hours after injury, but neither differences in stainingintensity nor the area encompassed by microglia were statisticallysignificant. After 5 days, both staining intensity and areas ofmicroglial activation were essentially the same for both groups ofanimals.

In addition, FIGS. 82A-82C illustrate some results discussed above. Inthe contusive study, animals were sacrificed and brains homogenized todetermine the EMF effect on the master pro-inflammatory cytokine. FIG.82A shows the results from the contusive study where EMF reduced IL-1βby approximately 10-fold in CSF in treated vs control animals.

In the invasive injury study, brains were collected in intact animals at0, 3.5, and 6 hours and assayed for levels of 1L-1β by ELISA. Resultsshown in FIG. 82B demonstrate that IL-1β levels in brain tissue werelower in injured rats treated with PEMF than that of the null group forboth models.

Similarly, FIG. 82C shows data from the same study where rats weresubjected to bilateral invasive penetrating needle injuries into thestriatum. CSF samples were collected under anesthesia from single ratsat time specified by the symbols shown in FIG. 82C and analyzed byELISA. The results suggest that IL-1β appears in CSF 6 hours afterinvasive trauma and, importantly, levels appear to be suppressed by PEMFtreatment.

These results indicate that EMF, configured according to embodimentsdescribed, produced a very rapid drop in the inflammatory response totraumatic brain and cervical injury which no other pharmacological orphysical modality has been able to achieve. An important factor is thatthese results were obtained with a portable disposable device which canbe incorporated in kits for field response to brain trauma, stroke andother neurological injuries.

Example 5

In this example, the effect of a radio frequency EMF signal, configuredaccording to an embodiment of the present invention consisting of a27.12 MHz carrier, pulse-modulated with a 3 msec burst repeating at 2 Hzand a peak amplitude of 0.05 G, on post-operative pain was studied in arandomized double-blind clinical study on breast reduction patients.Patients were treated with EMF, configured according to an embodiment ofthe present invention, delivered to the target tissue with a disposabledevice, similar to that illustrated in FIG. 78B, which was incorporatedin the post-surgical dressing.

Treatment regimen for active patients was 30 min every 4 hours for threedays. Sham patients received the same EMF device which did not deliver asignal. Wound exudates were collected and pain was assessed byparticipants using a validated Visual Analog Scale (VAS). Concentrationsof IL-1β, a major pro-inflammatory cytokine, were approximately 3-foldlower at 5 hours post-op (P<0.001) in wound exudates from EMF-treatedpatients compared to those of the control group. EMF also produced aconcomitant 2-fold decrease in pain at 1 hour (P<0.01) and a 2.5-folddecrease at 5 hours post-op (P<0.001), persisting to 48 hours post-op.No significant changes in VAS scores were observed in the control group.Furthermore, the increased levels of analgesia were reflected in a2.2-fold reduction in narcotic use in patients receiving activetreatment over the first 24 hours post-op (P=0.002). Importantly, thetime course for both pain and IL-1β reduction were concomitant, showingthat EMF, configured to modulate CaM/NO signaling in an embodimentaccording to the present invention, produced endogenous changes in thedynamics of IL-10 availability, which impacts the many known subsequentinflammatory events that are mediated by this cytokine, including thoseleading to post-operative pain. These results, which are illustrated inFIGS. 83A-B, demonstrate that EMF, configured according to an embodimentof the present invention produced a rapid, non-pharmacological,non-invasive post-operative anti-inflammatory response whichsignificantly reduced patient morbidity and the cost of health care, andenhanced healing.

Example 6

This example studies PEMF treatment to attenuate post-traumatic edema.PEMF signals, including a radiofrequency signals, have been shown toreduce the edema associated with various types of peripheral tissueinjury. For example, in a double-blinded study of human subjectsundergoing breast-reduction surgery, post-operative subjects weretreated with a PEMF signal consisting of a 27.12 MHz carrier,pulse-modulated with a 3 msec burst repeating at 2 Hz and a peakamplitude of 0.05 G. As shown in FIG. 85, wound exudates were collectedfor analysis and volumes were measured at regular post-operativeintervals. Results demonstrate a 30% reduction in volumes in the first 4hours after surgery. Asterisks in FIG. 85 indicate lower volumes in thegroup of post-operative subjects receiving PEMF treatment (*p<0.03).

With this current example, PEMF signals will be shown to attenuateincreases in brain volume, intracranial pressure, and T2-weighted MRIsignals. Animals will be subjected to the weight-drop injury andrandomly assigned to receive PEMF (or null) signals. Thirty rats will beimplanted with a Codman micro-sensor ICP probe (Codman, Raynham, Mass.)at the same time that the scalp is prepared for the weight-drop injury,as described for use in rats by Williams.

Using a stereotactic frame, a burr hole will be made at −4 mm posteriorto and 5 mm lateral to Bregma and the probe will be inserted to a depthof 2 mm. Baseline ICP will be monitored 10 minutes before the injury.The protruding part of the probe will be removed during impact. Afterinjury, 2 groups (n=15) will be treated with PEMF or null signals for 5minutes every 20 minutes for 8 hours and the sham group will bemaintained under similar conditions. The PEMF signal configuration usedmay be a sinusoidal wave at 27.12 MHz with peak magnetic field B=0.05 G(Earth=0.5 G), burst width, T1=5 msec, and repetition rate T2=2/sec asshown in FIG. 86A. The PEMF signal configuration may also induce a 1-5V/m peak electric field in situ with a duty cycle=2%, without heat orexcitable membrane activity produced. The field may be applied throughan electrical pulse generator to a coil tuned to 27.12 MHz. The burstwidth (5 msec) and repetition rate (2 Hz) were chosen by comparing thevoltage induced across the Ca²⁺ binding site over a broad frequencyrange to noise fluctuations over the same range. Effects of burst widthsof two 27.12 MHz sinusoidal signals at 1 Hz are illustrated in FIG. 86B.As shown in FIG. 86B, high signal-to-noise ratios (SNRs) can be achievedin the relatively low frequency range and at peak magnetic field 0.05 G.

Animals will be re-anesthetized at 30 minutes, 1 hour, 4 hours, and 8hours and the probe will be re-inserted for ICP measurement. After thefinal measurement, animals will be euthanized. ICP of both injury groupswill be compared over time with respect to pre- and post-injury valuesand effects of PEMF (vs. null) to determine the extent and kinetics ofICP for this model and to determine whether PEMF signals can attenuatethe magnitude of ICP or protract the rise in ICP over time.

T2-weighted Magnetic Resonance Imaging: Thirty rats will undergocontusive injuries and will be randomly assigned to receive PEMF or nullsignals (n=15) using a regimen of 5 minutes of treatment every 20minutes. T2-weighted MRI will be performed at the Gruss MagneticResonance Research Center (MRRC) at the Albert Einstein College ofMedicine, both before injury and after injury at 3 time points thatbracket peak ICP, as established in the pilot experiment (see above).Edema will be calculated using standard MRI algorithms and protocolsestablished at the MRRC. MRI is a validated method of following edema inpost injury neurotrauma models. Animals will be transported to the MRRCon a staggered basis. Under isofluorane anesthesia, each animal will beconnected to a ventilator and anesthesia will be maintained at 1.5%isofluorane. Ventilation rate will be maintained at 60 breaths/minute,and volume pressure settings will be adjusted to produce stableend-tidal CO2 and regular respiratory movements. Core temperature willbe monitored by rectal thermometer and a feedback-controlled water pumpwill warm the animal while in the MRI cradle. The animal will then beplaced into the magnet and imaging data are collected. The animal willthen be removed from the magnet, extubated, placed on afeedback-controlled warming pad, and allowed to recover from anesthesia,when it will be returned to its home cage and transported to the AnimalCare Facility.

Each T2W slice will be displayed on a workstation and edema will bequantified using the MEDx package after manually outlining areas ofsignal hyper-intensity that are consistent with edema. Volume will becomputed as the sum of area outlined on each slice multiplied by slicethickness. Longitudinal comparison and quantification of edema willallow values for each animal to be compared and normalized to its ownbaseline. Information from this analysis will include the determinationof areas of brain that are most affected by the injury and the abilityof PEMF to suppress brain swelling over the period of edema formation.Animals from this study will also be used for ¹H-MRS imaging.

ICP has been evaluated in response to severe injury in the weight-dropmodel. Normally, ICP ranges between 5 and 15 mm Hg. A 450 g weightdropped over 2 meters will result in a rise in ICP to 28±3 mm Hg after30 min, followed by a gradual decline, measured over 4 hours. Based onPEMF-mediated reductions of wound exudate volumes (FIG. 85), the studyresults are expected to show that PEMF to has immediate effects onreducing edema. Moreover, the ability to obtain whole brain images withT2-weighted MRI will allow us to identify regions of interest in ourmodel that incur the worst injury and follow them over time.

Example 7

In one study, a group of rats received neural transplants of dissociatedembryonic midbrain neurons and were treated twice a day with PEMF ornull signals for 1 week. As shown in FIG. 87, OX-42 labeled activatedmicroglia form a “cuff” surrounding the transplant. Alkalinephosphatase-labeled blood vessels were stained in purple. Results of themicroglial staining, shown in FIG. 87, demonstrate that microglialactivation was less intense in the PEMF group. This study showed thatPEMF may attenuate inflammation in response to transplantation. However,the apparent decrease in microglia may be transitory and that microglialactivity may actually be increased/accelerated.

Example 8

In this example, rats were subject to penetrating injuries and exposedto PEMF signals according to embodiments described. Brain tissue wasprocessed for OX-42 IHC at specified times after injury to identifyactivated microglia. As shown in FIG. 88, results demonstrate that thepattern of OX-42 staining in rats that received penetrating injuries waslocalized to the site of the trauma. Most importantly, stainingintensity appears higher with PEMF treatment at 2 and 5 days afterinjury, indicating activation of microglial cells was accelerated.

Example 9

As shown in FIG. 89, neuronal cultures were treated with PEMF signalsfor 6 days before challenge by (1) reduced serum 1% or (2) 5 μMquisqualic acid, a non-NMDA glutamate receptor agonist. Dopaminergicneurons were identified by tyrosine hydroxylase immunocytochemistry andquantified at 8 days. The bars shown in FIG. 89 indicate mean neuronalnumbers (+/−SEM) in triplicate cultures. Asterisk denotes groups withsignificant differences from the null group (P<0.05). Results indicatethat PEMF signals according to embodiments describe provideneuroprotective treatment to prevent neural death.

Example 10

This example will study the ability of PEMF signals to prevent neuraldeath. Animals will be subjected to contusive (weight-drop) TBI. Eightyrats will be randomly assigned to PEMF or null groups and treated for 5minutes every 20 minutes. A group of rats receiving sham surgery willserve as controls. At 1, 2, 5 and 10 days and after injury, CSF will becollected from 10 animals from each treatment group immediately prior toeuthanasia, at which time blood will be collected peri-mortem. Brainswill be fixed, cryoprotected, 50 μm vibratome sections will be generatedthrough the cerebrum from approximately −7 to +4 mm with respect toBregma on the anteroposterior axis. Multiple series of every 6th sectionwill be prepared for analyses described below.

Tissue Necrosis: The overall extent of tissue damage will be assessed ona series of sections after hematoxylin and eosin (H&E) histochemistry,first qualitatively, by observations of astrocytic, neuronal, ordendritic swelling, pyknotic nuclei, and necrosis and thenquantitatively, by measuring the area abnormal histology. Regions ofdamaged tissue will be captured by digital photography and the volumewill be assessed by outlining the perimeter using Image J software,calculating the area with calibrated markers, and multiplying by sectionthickness. Histological abnormalities, as described above, will bequantified within a specified volume of these regions. Data from eachgroup will be compared to determine whether PEMF signals reduce thevolume of tissue damage and numbers of cells with abnormal morphology.

Neuronal Injury: Proton Magnetic Resonance Spectroscopy: This study willbe conducted on the same animals that will be used for T2-MRI studies(see Example 6), as they will provide information on regions ofinterest, and to avoid duplications of time and costs associated withlive-animal studies. Based on information obtained from T2-weighted MRI(see Example 6), several regions of interest (ROI), defined byanatomical landmarks and changes on T2 maps, will be selected andfurther analyzed. Studies by other groups suggest that both cortical andhippocampal regions may be the most vulnerable to the injury made in theweight-drop model. Afterwards, computerized graphical analysis ofspecific, localized spectra in the ROIs will be utilized to determineresonance corresponding to NAA, Cr, Cho, lactate and taurine.Quantitative analysis of the spectroscopic metabolite ratios will becompared among the pre-injury, TBI null, and TBI PEMF groups todetermine changes in concentrations of these biochemical markers.

Neuronal Death: Fluoro-Jade staining: Fluoro-jade stain is afluorochrome derived from fluorescein and is commonly used to labeldegenerating neurons including neurons injured from TBI as analternative to other methods, such as silver and Nissl stains.Fluoro-jade stained tissue can be visualized with epifluorescence usingfilters designed for fluorescein or fluorescein isothiocynate(excitation 495 nm; emission 521 nm). Multiple morphological featurescan be detected using fluoro-jade including; cell bodies, dendrites,axons, and axon terminals. Even though all fluoro-jade derivatives candetect these specific morphological features, fluoro-jade C has greaterspecificity and resolution. A series of every 6th serial section will beprocessed stained with fluoro-jade-C to identify dying neurons. Sectionswill be dehydrated in ethanol and rinsed in distilled water, followed byoxidation in 0.06% potassium permanganate for 15 minutes, followed byseveral water washes. Sections will then be placed in 0.1% acetic acidcontaining 0.001% fluoro-jade C (Millipore) for 1 hour at roomtemperature. After washing, sections will be dehydrated, cleared andcoverslipped for viewing. Areas demonstrating the greatest generalizeddamage by H&E will be assessed for neuronal damage. Neurons within adefined anatomical structure can be quantified on a series of sectionsby stereological analysis (i.e. optical dissector) using Neurolucidasoftware (Microbrightfield).

Data from PEMF-treated and null groups will be compared. UCH-L1:Ubiquitin C-terminal hydrolase-L1 (UCH-L1), a neuron-specific protein(also called protein gene product 9.5 or Park 5) involved in proteindegradation via the ATP-dependent proteosomal pathway, is abundant inneuronal cell bodies. Mice bearing a spontaneous mutation in this genedemonstrate behavioral disturbances and neuronal loss, and mutations inhumans are associated with Parkinson's disease, supporting a sustainingrole for this protein in neurons. Importantly, UCL-L1 was identified ina proteomic screen of CSF as a biomarker for neuronal injury. Studieshave shown that UCH-L1 is released following severe cortical impactinjury and ischemia. This marker has recently gained attention in thegeneral press as a potential CSF marker for brain injury in humans.Therefore, we will evaluate the effects of PEMF on UCH-L1 in CSF, and wewill also assess levels in blood, as they can easily be obtained beforeeuthanasia. Although not commercially available, we will devise an ELISAto quantify UCH-L1 using chicken and rabbit UCH-L1 polyclonal antibodies(Cell Signaling and Thermo Scientific, respectively), as described byothers. Plates will be coated with anti-UCH-LI, followed by washing andaliquots of CSF, or blood. Protein will be identified with HRP-antiUCH-L1 and a soluble substrate for peroxidase. Western blots for UCH-L1(a 24 kD protein) will be run with selected samples to validate ELISAdata. Levels of UCH-L1 should be inversely proportional to the extent ofneuronal death.

Axonal Injury: A series of sections from injured animals in PEMF andnull groups and sham controls will be processed for silver staining withthis method. Briefly, mounted tissue sections are pre-incubated in analcoholic solution containing silver and copper nitrates, washed inacetone, and impregnated in silver nitrate with lithium and ammoniumhydroxides, followed by reduction in formalin, citric acid and ethanol.After acidification, bleaching, and fixation, slides will becoverslipped for viewing. Tissue sections will be processed commercially(Neuroscience Associates, Knoxville, Tenn.), as this technique requiresa number of hazardous solutions that require special processing anddisposal. Silver-impregnated, degenerating neurons and processes willstain black, progressing to a Golgi-like intensity. More lightly stainedterminals and lysosomes may only be apparent at earlier time points, asthese structures often degenerate prior to axonal loss. Forquantification of degeneration, images will be digitized and the densityof optical staining over an assigned area of cortex will be quantifiedby densitometry with Image J Software. This method has been validated byothers.

Predicted Results: The time course of pathological events following TBIare the direct destruction of tissue (including neurons) if the injuryis invasive, followed by edema, inflammation, axonal injury, andsubsequently delayed neuronal death. The Marmarou weight-drop methodinduces all of these events in a more protracted fashion. Cell cultureexperiments indicate that neuronal survival is increased with PEMFdirectly in response to an excitotoxic insult, suggesting thatneurotoxicity due to ischemia and subsequent release of glutamate mayalso be attenuated by PEMF signals in vivo. Because brain swelling andinflammation result in indirect neurotoxicity, increased survival byPEMF is also predicted for this pathway. Positive results will confirmthat treatment with PEMF signals can be used to attenuate the damagecaused by traumatic, closed head injury and may have therapeuticimplications for other types of TBI as well as more acute and chronicneurodegenerative diseases, such as stroke, Alzheimer's disease, andParkinson's disease where many of these same mechanisms are known to beinvolved.

Example 11

In this example, rats will be subjected to the Marmarou weight-drop TBImodel to produce moderate behavioral deficits. Individual naive animalswill be subjected to general assessments and sensorimotor behavioraltesting. Those animals whose behavior falls in the normal range willreceive moderate TBI using the weight-drop model and will be randomlyassigned to receive PEMF or null signals using the regimen of 5 minutesevery 20 minutes. At 1, 2, 7, 14, and 21 days after injury, animals willbe re-assessed for general behaviors and neurological function toquantify the magnitude of these basic deficits. At 1 month aftersurgery, animals will be transported to the Bronx VA for long-termcognitive testing. After acclimation to the VA animal holding facility(2 weeks), testing will take place over 8 weeks for each animal by thesame technician.

Rats will first be evaluated for general health and spontaneous andelicited behavior. These basic observations will be supplemented with anassessment of motor, sensory and general activity level using rotarod,grip strength, balance beam, and tail-flick analgesia tasks to determinewhether the injury has affected the general health status and overtbehavioral profile of the rat in a way that would make its generalbehavior incompatible with more complex behavioral assays. Moreover, ifspecific deficits are identified in the basic screen, we may be able toalter the choice of more complex behavioral assays to account for thedeficit. We will then proceed to more detailed testing. Rats will beobserved in an open field assay to assess both general motor activityand anxiety related behavior, and an elevated plus maze as an additionalindicator of anxiety related behavior. In the cognitive domain we willadminister at least three tests designed to measure learning and memoryrelated functions: 1) the Morris water maze, a standard test ofhippocampal dependent spatial memory 2) a test of contextual and cuedfear conditioning, which is highly dependent on amygdaloid function andrequires a set of motor and sensory abilities distinct from thoserequired for spatial navigation, and 3) a Y-maze task, a test of workingmemory. We will also measure response to acoustic startle and pre-pulseinhibition as measures of auditory function and sensory gating,physiological functions that can be affected in TBI.

Order of testing and timetable: Carryover effects can significantlyconfound behavioral testing in rodents. The testing order will be asdescribed except that the cued fear conditioning and Morris water mazetests will be performed last in the sequence, as these include the mostdemanding and stressful tasks. Based on our prior experience in rodentbehavioral work, testing will require: Basic screen (SHIRPA) (7 days),Rotarod (2 days), Grip strength (1 day), Tail flick (1 day), open field(3 days), elevated plus maze (2 days), Morris water maze (4 weeks),contextual/cued fear conditioning (2 days), Y-maze (2 days) and acousticstartle/PPI (2 days) or approximately 8 weeks of testing.

Data analysis: Data will be analyzed using GraphPad Prism 5.0 (GraphPadSoftware, San Diego, Calif.) or SPSS 18.0 (SPSS, Chicago, Ill.) softwareas in previous studies. Depending on the behavioral test, statisticswill employ univariate or repeated measures analysis of variance(ANOVA), unpaired t-tests or linear regression. Equality of variancewill be assessed using the Levene test and when it is not significant(p>0.05) between-group comparisons will be made with unpaired t-tests(Student's) or Tukey post-hoc tests. If the Levene statistic issignificant (p<0.05) unpaired t-tests will be used using the Welchcorrection for unequal variances. For repeated-measures ANOVA,sphericity will be assessed using Mauchly's test. If the assumption ofsphericity is violated (p<0.05, Mauchly's test), significance will bedetermined using the Greenhouse-Geisser correction.

Predicted Results: Data from PEMF and null groups will be compared withnaive animals to determine the degree of deficit and with each other todetermine whether PEMF signals improve neurological function. It isexpected that PEMF treatment will show a decrease in the degree ofinitial deficits and/or accelerate or enhance the degree of recovery.

As for additional details pertinent to the present invention, materialsand manufacturing techniques may be employed as within the level ofthose with skill in the relevant art. The same may hold true withrespect to method-based aspects of the invention in terms of additionalacts commonly or logically employed. Also, it is contemplated that anyoptional feature of the inventive variations described may be set forthand claimed independently, or in combination with any one or more of thefeatures described herein. Likewise, reference to a singular item,includes the possibility that there are plural of the same itemspresent. More specifically, as used herein and in the appended claims,the singular forms “a,” “and,” “said,” and “the” include pluralreferents unless the context clearly dictates otherwise. It is furthernoted that the claims may be drafted to exclude any optional element. Assuch, this statement is intended to serve as antecedent basis for use ofsuch exclusive terminology as “solely,” “only” and the like inconnection with the recitation of claim elements, or use of a “negative”limitation. Unless defined otherwise herein, all technical andscientific terms used herein have the same meaning as commonlyunderstood by one of ordinary skill in the art to which this inventionbelongs. The breadth of the present invention is not to be limited bythe subject specification, but rather only by the plain meaning of theclaim terms employed.

While the apparatus and method have been described in terms of what arepresently considered to be the most practical and preferred embodiments,it is to be understood that the disclosure need not be limited to thedisclosed embodiments. It is intended to cover various modifications andsimilar arrangements included within the spirit and scope of the claims,the scope of which should be accorded the broadest interpretation so asto encompass all such modifications and similar structures. The presentdisclosure includes any and all embodiments of the following claims.

What is claimed is:
 1. A method for treating a neurological injury orcondition caused by a stroke in a patient in need thereof, the methodcomprising: generating a pulsed electromagnetic field from a pulsedelectromagnetic field source; and applying the pulsed electromagneticfield in proximity to a target region affected by the neurologicalinjury or condition caused by the stroke to reduce a physiologicalresponse to the neurological injury or condition.